Atlas of FSHR expression from novel reporter mice

Research facility: Shanxi Medical University

Published Mar 17, 2025 on Dryad. https://doi.org/10.5061/dryad.1jwstqk4t

Abstract

The FSH-FSHR signaling pathway has traditionally been considered an essential regulator in reproductive development and fertility. But there has been emerging evidence of FSHR expression in extragonadal tissues/organs. This poses new questions and long-term debates regarding the physiological role of the FSH-FSHR pathway, and underscores the need for reliable, in vivo analysis of FSHR expression in animal models. However, conventional methods have proven insufficient for examining FSHR expression due to limitations, such as the scarcity of ‘reliable’ antibodies, rapid turnover/degradation of transcripts, and a lack of robust in vivo tools. To address this challenge, we developed Fshr-ZsGreen ‘knockin’ reporter mice under the control of Fshr endogenous promoter using CRISPR/Cas9 genome-editing technology to append a P2A-ZsGreen targeting vector into a locus between the last exon and the stop codon of Fshr. With this novel genetic tool, we provide a reliable readout of Fshr expression at single-cell resolution level in vivo and in real time. Reporter animals were also subjected to additional analyses, including immunohistochemical staining, ddRT-PCR, and in situ hybridization, to define the accurate expression profile of FSHR in gonadal and extragonadal organs/tissues. Our compelling results not only demonstrated Fshr expression in intragonadal tissues but also, strikingly, unveiled notably increased expression in Leydig cells, osteoblast lineage cells, endothelial cells in vascular structures, and epithelial cells in bronchi of the lung and renal tubes. The genetic decoding of the widespread pattern of Fshr expression highlights its physiological relevance beyond reproduction and fertility, and opens new avenues for therapeutic options for age-related disorders of the bones, lungs, kidneys, and hearts, among other tissues/organs. Exploiting the power of the Fshr knockin reporter animals, this report provides the first comprehensive genetic record of the spatial distribution of FSHR expression, correcting a long-term misconception about Fshr expression and offering prospects for extensive exploration of FSH-FSHR biology.

https://doi.org/10.5061/dryad.1jwstqk4t

Description of the data and file structure

This dataset includes raw data for gel images and microscope images for data published in our manuscript at the eLife, which are compressed and can be depressed with ZIP.

The submitted files are in two forms:

1) *TIFF images can be viewed by Photoshop or Image J;

2) *xlsx files can open the Eslsx file.

The description of these data:

1. Fig. 1 B and C images for genotyping PCR and Southern blots: 2-sub-folders

(1) Fig. 1B PCR images for genotyping-4 images in TIF Fig. 1B-b and c

Two primer pairs were used for genotyping heterozygote mice to produce the following two images:

1) Uncropped image for Fig. 1B-b;

2) Uncropped image for Fig. 1B-c;

3) Labelled images for Figure1B-b;

4) Labelled images for Figure1B-c.

(2) Fig. 1C images for Southern blots-6 TIFF images

Two probes were used for Southern blots: 5’ (SCal I) and 3’ (Ncol) probes for the following images:

1) Uncropp original image for Fig, 1B-c left 1 using a 5’ probe;

2) Uncropp original image for Fig, 1B-c left 2 using a 5’ probe;

3)) Uncropp original image for Fig, 1B-c right 1 using a 3’ probe;

4) Uncropp original image for Fig, 1B-c right 2 using a 3’ probe;

5) labelled Original images for Figure 1-C-b right using 5’ probe;

6) labelled Original images for Figure 1-C-b left using 3’ probe.

2. Fig. 2A Ovary: 8 images in TIFF, showing Fshr co-expression with Stra8.

1) Fig. 2A-a Fshr-ZsG with Stra8 _10X _RGB;

2) Fig. 2A-b Fshr-ZsG with Stra8 _20X _RGB;

3) Fig. 2A-c Fshr-ZsG with Stra8 _20X _RGB;

4) Fig. 2A-d Fshr-ZsG with Stra8 _20X _RGB;

5) Fig. 2A-e Fshr-ZsG with Stra8 100X_RGB;

6) Fig. 2A-f Fshr-ZsG with Stra8 100X_RGB;

7) Fig. 2A-g Fshr-ZsG with Stra8 100X_RGB;

8) Fig. 2A-h Fshr-ZsG with Stra8 100X_RGB.

3. Fig. 2B C D E F and G Testis:

(1) Fig. 2B Fshr co-expression with Stra8: 3 images in TIFF

a. Fig. 2B-a Fshr-ZsG with Stra8 4X;

b. Fig. 2B-b Fshr-ZsG with Stra8 40X_RGB;

c. Fig. 2B-c Fshr-ZsG with Stra8 100X_RGB.

(2) Fig. 2B-Fshr co-expression with SET: 3 images in TIFF

d. Fig. 2B-d Fshr-ZsG with Stra8 40X_RGB;

e. Fig. 2B-e Fshr-ZsG with Set 100X_RGB;

f. Fig. 2B-f Fshr-ZsG with Set 100X_RGB- showing a vascular tube.

(3) Fig. 2C-RNA-smFISH confirmation of Fshr expression in Fshr-ZsG mice: 2 folders

1) Fig. 2C-Fshr sense probe: 2 sub-folders

a. 40X: 4 images in TIFF-showing 3 channels with one overlap;

b. 100X: 4 images in TIFF–showing 3 channels with one overlap.

2) Fig.2C-Fshr antisense probes:

a. 40X: 4 images in TIFF-showing 3 channels with one overlap;

b. 100X: 4 images in TIFF-showing 3 channels with one overlap.

(4) Fig. 2D-Fshr-smFISH images in B6: 6 images in TIFF

1) Fig. 2D-a B6 Testis negative control sense probe 40X_RGB;

2) Fig. 2D-b B6 Testis negative control sense probe 40X_RGB;

3) Fig. 2D-c B6 Testis negative control sense probe 40X_RGB;

4) Fig. 2D-d B6 Testis Fshr antisense probe40X_RGB;

5) Fig. 2D-e B6 Testis Fshr antisense probe40X_RGB;

6) Fig. 2D-f B6 Testis Fshr antisense probe40X_RGB.

(5) Fig. 2E Fshr in B6: 2 sub foldersFig. 2E-a

1) Fshr in the ovary: 4 images in TIFF

a. Fig. 2E B6 ovary IgG 4X_RGB;

b. Fig. 2E B6 ovary IgG 40X_RGB;

c. Fig. 2E B6 ovary Fshr Ab.-4X_RGB;

d. Fig. 2E B6 ovary Fshr Ab.-40X_RGB

2) Fshr in the testis: 4 images in TIFF

a. Fig. 2E B6 testis IgG 4X_RGB;

b. Fig. 2E B6 testis IgG 40X_RGB;

c. Fig. 2E B6 testis Fshr Ab.-4X_RGB;

d. Fig. 2E B6 testis Fshr Ab.-40X_RGB

(6) Fig. 2F-Fshr in TM3 cells-8 images in TIFF

1) Fig. 2F-a TM3 cells isotype IgG 40X_RGB;

2) Fig. 2F-b TM3 cells Fshr Ab. 40X_RGB;

3) Fig. 2F-c TM3 cells Fshr Ab. 100X_RGB;

4) Fig. 2F-d TM3 cells Fshr Ab. single layer 100X_RGB;

5) Fig. 2F-e TM3 cells Fshr Ab. 3D 100X_RGB;

6) Fig. 2F-f TM3 cells Fshr Ab. nuclei 100X_RGB

7) Fig. 2F-g TM3 cells Fshr in the nucei single layer _RGB;

8) Fig. 2F-h TM3 cells Fshr in the nucei SIM 3D 100X _RGB.

(7) Fig. 2G Fshr expression by dd.rtPCR: one Excel file.

4. Fig. 3 Skeletal tissues: 23 TIFF images

(1) Fig. 3A-a Fshr-ZsG in growth plate: 1 image in TIFF;

(2) Fig. 3A-b Fshr-ZsG in epiphyseal area: 1 image in TIFF;

(3) Fig. 3A-c Fshr-ZsG in trabecular bone: 1 image in TIFFl

(4) Fig. 3A-d Fshr-ZsG in the endosteal area: 1 image in TIFF;

(5) Fig. 3A-e Fshr-ZsG in chondryocytes: 1 image in TIFF;

(6) Fig. 3A-f Fshe-ZsG in Ostteoblasts and osteoclasts: 1 image in TIFF;

(7) Fig. 3A-g Fshr-ZsG in trabecullar bone: 1 image in TIFF;

(8) Fig. 3A-h Fshr-ZsG in cells on the endosteal surface: 1 image in TIFF;

(9) Fig. 3B-a Fshr and osteocalcin in chondrycotes: 1 image in TIFF;

(10) Fig. 3B-b Fshr-ZsG with osteocalcin in Obs: 1 image in TIFF;

(11) Fig. 3B-c Fshr-ZsG and osteocalcin bone lining cells: 1 image in TIFF;

(12) Fig. 3B-d Fshr-ZsG and osteocalcin in the cortex: 1 image in TIFF;

(13) Fig. 3B-e Fshr-ZsG and Trap in osteoclasts: 1 images in TIFF;

(14) Fig. 3B-f Fshr-ZsG and Trap in Obs and bone lining cells

(15) Fig. 3C-a-FshrZsG and CD34 in BM: 1 image in TIFF;

(16) Fig. 3C-b Fshr-ZsG and CD34 condryocytes: 1 image in TIFF;

(17) Fig. 3C-c Fshr-ZsG and CD133 in BM: 1 image in TIFFl

(18) Fig. 3C-d Fshr-ZsG and CD133 in chondryocytes: 1 image in TIFF;

(19) Fig. 3C-e Fshr-ZsG and CD133 in the cortex: 1 image in TIFF;

(20) Fig. 3D-Control Cortex 40x_RGB;

(21) Fig. 3D Fshr cKO Cortex 40x_RGB;

(22) Fig. 3D Control Trabecular bone 40x_RGB;

(23) Fig. 3D Fshr cKO Trabecular bone 40x_RGB.

5. Fig. 4 Fshr-ZsG in WAT and BAT: 8 sub-folders

(1) Fig. 4A 4X: 1 image in TIFF showing one piece of WAT;

(2) Fig. 4A a, b and c 40X: 3 TIFF images of Fshr-ZsG expression from three different areas of Fig. 4A 4X;

(3) Fig. 4B Fshr-ZsG expression with Ucp1: 4 TIFF images of Fshr-ZsG expression from different areas of WAT;

(4) Fig. 4C Fshr-ZsG in BAT: one 4X image and three 40X images showing Fshr-ZsG expression in 3 different areas of 4X image;

(5) Fig. 4D BAT Fshr-ZsG and Ucp1: 1-4X image, 3-40X images and 3-100X images-showing Fshr-ZsG expression with Ucp1 in BAT in different areas;

(6) Fig. 4E BAT Fshr-ZsG and TH: 1-4X image, 3-40X images and 3-100X images-showing Fshr-ZsG expression with TH in BAT in different areas;

(7) Fig. 4F BAT Fshr-ZsG and Peri: 1-4X image, 3-40X images and 3-100X images-showing Fshr-ZsG expression with Peri in BAT in different areas;

(8) Fig. 4G WAT and BAT with Fshr Ab in B6: showing Fshr expression in different areas of WAT and BAT from B6 mice. 2 sub-folders:

1) Fig. 4G BAT Ab: 2-4X TIFF images and 2-40X TIFF images for control and antibody treated, respectively;

2) Fig. 4G WAT Fshr Ab: 2-4X TIFF and 2-40X TIFF images for control and antibody treated, respectively.

6. Fig. 5 Heart and Aorta: 3 sub-folders

(1) Fig. 5A: two TIFF images-

a. Fig. 5A-a cardiomyocytes 40X _RGB;

b. Fig. 5A-b smooth muscles 40X _RGB.

(2) Fig. 5B: Fshr-ZsG co-expression with a-SMA. 14 TIFF images:

1) Fig. 5B-Whole heart 4X _RGB;

2) Fig. 5B-a cross-oriented cardiomyocytes 40X _RGB;

3) Fig. 5B-b longitudinally oriented cardiomyocytes 40X _RGB;

4) Fig. 5B-c cross-oriented smooth muscle of the ascending aorta with brown adipose tissue 40X _RGB;

5) Fig. 5B-d smooth muscle of the left pulmonary artery with BAT 40X_RGB;

6) Fig. 5B-e BAT attached to a large artery 40X _RGB;

7) Fig. 5B-f a layer of endothelial cells of the superior vena cava 40X _RGB;

8) Fig. 5B-g connective tissue between arteries with BAT and nerves 40X _RGB;

9) Fig. 5B-h SVC with smooth muscle and connective 40X_RGB;

10) Fig. 5B-i cardiomyocytes with an arteriole 100X_RGB;

11) Fig. 5B-j BAT attached to the large artery 100X_RGB;

12) Fig. 5B-k endothelial cells 100X_RGB

13) Fig. 5B-m transverse section of nerve fibers 100X_RGB;

14) Fig. 5B-n the layers of ECs and SM of the SVC. 100X_RGB.

(3) Fig. 5C Fshr-ZsG and EMCN expression in the heart: 5 TIFF images

(1) Fig. 5C whole heart Fshr-ZsG and EMCN 4X_RGB;

(2) Fig. 5C-a SM and ECs Fshr-ZsG and EMCN 40X-RGB;

(3) Fig. 5C-b pulmonary artery Fshr-ZsG and EMCN 100X_RGB;

(4) Fig. 5C-c cardiomyctes 100X_RGB;

(5) Fig. 5C-d ECs 100X_RGB.

(4) Fig. 5D: 6 TIFF images with Fshr-ZsG and a-SMA and EMCN expression in the Aorta

1) Fig. 5D a longitudinal sectioned aorta with Fshr-ZsG and a-SMA 10X _RGB;

2) Fig. 5D-b aorta with Fshr-ZsG and a-SMA 100X _RGB;

3) Fig.. 5D-c aorta with Fshr-ZsG and EMCN 100X_RGB;

4) Fig. 5D-d transversely sectioned aorta with Fshr-ZsG 10X-RGB;

5) Fig. 5D-e transversely sectioned aorta with Fshr-ZsG and a-SMA 100X-RGB;

6) Fig. 5D-f transversely sectioned aorta with Fshr-ZsG and EMCN 100X-RGB

7. Fig. 6A Lung: 9 TIFF images of lung section with Fshr-ZsG and PDL-1 staining

1) Fig. 6A - Left a large areas of lung with Fshr-ZsG and PDL-1 4X _RGB;

2) Fig. 6A -a ciliated columnar cells of primary bronchi 40X _RGB;

3) Fig. 6A -b bronchioles with alveoli 40X _RGB;

4) Fig. 6A -c respiratory bronchiole with bronchial gland 40X _RGB;

5) Fig. 6A -d alveoli 40X _RGB;

6) Fig. 6A -e ciliated columnar cells of primary bronchi 100X _RGB;

7) Fig. 6A -f the bronchioles with alveoli 100X _RGB;

8) Fig. 6A -g respiratory bronchiole with bronchial gland 1000X _RGB;

9) Fig. 6A -h alveoli 100X _RGB.

8. Fig. 6B Kidney: 8 TIFF images of lung section with Fshr-ZsG and Col1a1 staining

1) Fig. 6B-a whole kidney 4X _DAPI_RGB_blue;

2) Fig. 6B-b whole kidney Col1a1 4X _RGB_red;

3) Fig. 6B-c whole kidney Fshr-ZsG 4X_RGB _green;

4) Fig. 6B-d whole kidney overlap 4X _RGB;

5) Fig. 6B-e proximal glomerulus and renal tubes 40X _RGB;

6) Fig. 6B-f distal glomerulus and renal tubes 40X _RGB;

7) Fig. 6B-g glomerulus 100X _RGB;

8) Fig. 6B-h arteriole 100X _RGB.

9. Fig. 7 Liver: 3 sub-folders

1) Fig. 7A Fshr-ZsG and Col1a1 staining:

a. Fig. 7A-a hepatic cells with a central vein Fshr-ZsG and Col1a1 40X_RGB;

b. Fig. 7A-b hepatic artery Fshr-ZsG and Col1a1 40X _RGB;

c. Fig. 7A-c hepatic cells with a central vein Fshr-ZsG and Col1a1 100X_RGB;

d. Fig. 7A-d hepatic artery Fshr-ZsG and Col1a1 100X _RGB.

2) Fig. 7B Fshr-ZsG and CD31 staining

a. Fig. 7B-a heptaocytes Fshhr-ZsG and CD31 40X _RGB;

b. Fig. 7B-b heptaocytes Fshhr-ZsG and CD31 100X _RGB.

3) Fig. 7C Fshr-ZsG and KCNM1 staining

a. Fig. 7C-a heptaocytes ith a CV KCNMA1 40X _RGB;

b. Fig. 7C-b heptaocytes ith a CV KCNMA1 100X _RGB;

c. Fig. 7C-c hepatic cells with peripheral nerve fibers KCNMA1 40X _RGB;

d. Fig. 7C-d hepatic cells with peripheral nerve fibers KCNMA1 100X _RGB;

e. Fig. 7C-e small nerve fibers located around a vein KCNMA1 40X _RGB;

f. Fig. 7C-f small nerve fibers located around a vein KCNMA1 100X _RGB.

10. Fig. 8 Pancreas: 3 sub-folders

1) Fig. 8A pancreas_Fshr-ZsG and NG3 staining

a. Fig. 8A Pancreas_ Fshr-ZsG and NG3 40X_RGB;

b. Fig. 8A Pancreas_ Fshr-ZsG and NG3 100X_RGB.

2) Fig. 8A pancreas_Fshr and Insulin staining

a. Fig. 8B Pancreas_ Fshr-ZsG and Insulin 40X_RGB;

b. Fig. 8B Pancreas_ Fshr-ZsG and Insulin 100X_RGB.

3) Fig. 8A pancreas_Fshr and Glucogen staining

a. Fig. 8B Pancreas_ Fshr-ZsG and Glucogon 40X_RGB;

b. Fig. 8B ancreas_ Fshr-ZsG and Glucogon 100X_RGB.

11. Fig. 9 Thyroid: 5 TIFF images

a. Fig. 9-Left panel Fshr-ZsG with Tshr 4X_RGB;

b. Fig. 9a-The edge of 4X thyroid Fshr-ZsG with Tshr 40X_RGB;

c. Fig. 9b-The edge of 4X thyroid Fshr-ZsGwith Tshr 40X_RGB;

d. Fig. 9c-The center of thyroid Fshr-ZsGwith Tshr 100X_RGB;

e. Fig. 9d-The center of thyroid Fshr-ZsGwith Tshr 100X_RGB.

12. Fig. 10 Skin: 2 sub-Folders

1) Fig. 10A Fshr-ZsG: 2 TIFF images

a. Fig. 10A-a Longitudinal section of skin Fshr-ZsG 40X _RGB;

b. Fig. 10A-b Transverse section of skin Fshr 40X_RGB.

2) Fig. 10B Fshr-ZsG and CD34: 2 TIFF images

a. Fig. 10A-a hair follicles Fshr-ZsG and CD34 staining 40X _RGB;

b. Fig. 10A-b hair follicles Fshr-ZsG and CD34 staining 100X _RGB.

13. Fig. 11 Skeletal muscles: 9 TIFF images

a. Fig. 11-a skeletal muscle Fshr with a-SMA 4X_RGB;

b. Fig. 11-b longitudinal section of skeletal muscle Fshr with a-SMA 40X_RGB;

c. Fig. 11-c longitudinal section of skeletal muscle Fshr with a-SMA 100X_RGB;

d. Fig. 11-d cross-section of skeletal muscle Fshr with a-SMA 40X_RGB;

e. Fig. 11-e cross-section of skeletal muscle Fshr with a-SMA 100X_RGB;

f. Fig. 11-f longitudinal section of skeletal muscle Fshr with PAX7 40X_RGB;

g. Fig. 11-g longitudinal section of skeletal muscle Fshr with PAX7 100X_RGB;

h. Fig. 11-h cross-section of skeletal muscle Fshr with TH 40X_RGB;

i. Fig. 11-i cross-section of skeletal muscle Fshr with TH 100X_RGB.

14. Fig. Spleen and bone marrow: 3-sub-folders

1) Fig. 12A Spleen Fshr-ZsG and CD11b staining

a. Fig. 12A- Left panel whole spleen Fshr-ZG with CD11b staining 4X_RGB;

b. Fig. 12A-a an area located at the edge of the spleen Fshr-ZsG with CD11b 40X_RGB;

c. Fig. 12A-b an area located at the edge of the spleen Fshr-ZsG with CD11b 100X_RGB;

d. Fig. 12A-c an area of red pulp Fshr-ZsG with CD11b 40X_RGB

e. Fig. 12A-d an area of red pulp Fshr-ZsG with CD11b 100X_RGB;

f. Fig. 12A-e an area of white pulp Fshr-ZsG with CD11b 40X_RGB;

g. Fig. 12A-f an area of white pulp Fshr-ZsG with CD11b 100X_RGB.

2) Fig. 12B: 4 TIFF images for Fshr-ZsG and CD3 staining

a. Fig. 12B-a whole spleen Fshr with CD3 4X_RGB;

b. Fig. 12B-b Trabecule of the spleen Fshr with CD3 40X_RGB;

c. Fig. 12B-c white pulp of spleen Fshr with CD3 100X_RGB;

d. Fig. 12B-d red pulp of spleen Fshr with CD3 100X_RGB.

3) Fig. 12C 2 TIFF images of BM for Fshr-ZsG and F4/89 staining

a. Fig. 12C-a Fshr with F4/80 the center of BM 100X_RGB;

b. Fig. 12C-b Fshr with F4/80 BM close to the cortex 100X_RGB.

15. Fig. 13 Brain: 9 TIFF images for Fshr-ZsG and staining of GFAP, Iba1 and NeuN

a. Fig. 13-a Olfactory bulb Fshr with GFAP 100X_RGB;

b. Fig. 13-b Pallidum Fshr with GFAP 100X_RGB;

c. Fig. 13-c Hippocampus Fshr with GFAP 100X_RGB;

d. Fig. 13-d Olfactory bulb Fshr with Iba1 100X_RGB;

e. Fig. 13-e Pallidum Fshr with Iba1 100X_RGB;

f. Fig. 13-f Hippocampus Fshr with Iba1 100X_RGB;

g. Fig. 13-g Olfactory bulb Fshr with NeuN 100X_RGB;

h. Fig. 13-h Pallidum Fshr with NeuN 100X_RGB;

i. Fig. 13-i Hippocampus Fshr with NeuN 100X_RGB.

16. Fig. 14A Confirmation of Fshr-ZsG expression in mouse organs by Fshr Ab.: 10 sub-folders

1) Fig. 14A Aorta-4 TIFF images

a. Fig. 14A-Aorta DAPI 100X_RGB_blue;

b. Fig. 14A-Aorta Ab 100X_RGB_red;

c. Fig. 14A-Aorta GFP 100X_RGB_green;

d. Fig. 14A-Aorta overlap 100X_RGB.

2) Fig. 14A Cardiac muscle

a. Fig. 14A-Cardiac muscle DAPI 100X_RGB_blue;

b. Fig. 14A-Cardiac muscle Ab 100X_RGB_red;

c. Fig. 14A-Cardiac muscle GFP 100X_RGB_green;

d. Fig. 14A-Cardiac muscle overlap 100X_RGB.

3) Fig.14A Kidney

a. Fig. 14A-Kidney DAPI 100X_RGB_blue;

b. Fig. 14A-Kidney Ab 100X_RGB_red;

c. Fig. 14A-Kidney GFP 100X_RGB_green;

d. Fig. 14A-Kidney overlap 100X_RGB.

4) Fig. 14A Liver

a. Fig. 14A-Liver DAPI 100X_RGB_blue;

b. Fig. 14A-Liver Ab 100X_RGB_red;

c. Fig. 14A-Liver GFP 100X_RGB_green;

d. Fig. 14A-Liver overlap 100X_RGB.

5) Fig. 14A Lung

a. Fig. 14A-Lung DAPI 100X_RGB_blue;

b. Fig. 14A-Lung Ab 100X_RGB_red;

c. Fig. 14A-Lung GFP 100X_RGB_green;

d. Fig. 14A-Lung overlap 100X_RGB.

6) Fig. 14A BAT

a. Fig. 14A-BAT DAPI 100X_RGB_blue;

b. Fig. 14A-BAT Ab 100X_RGB_red;

c. Fig. 14A-BAT GFP 100X_RGB_green;

d. Fig. 14A-BAT overlap 100X_RGB.

7) Fig. 14A Bone

a. Fig. 14A-Bone DAPI 100X_RGB_blue;

b. Fig. 14A-Bone Ab 100X_RGB_red;

c. Fig. 14A-Bone GFP 100X_RGB_green;

d. Fig. 14A-Bone overlap 100X_RGB.

8) Fig. 14A Ovary

a. Fig. 14A-Ovary DAPI 100X_RGB_blue;

b. Fig. 14A-Ovary Ab 100X_RGB_red;

c. Fig. 14A-Ovary GFP 100X_RGB_green;

d. Fig. 14A-Ovary overlap 100X_RGB.

9) Fig. 14A Thyroid

a. Fig. 14A-Thyroid DAPI 100X_RGB_blue;

b. Fig. 14A-Thyroid Ab 100X_RGB_red;

c. Fig. 14A-Thyroid GFP 100X_RGB_green;

d. Fig. 14A-Thyroid overlap 100X_RGB.

10) Fig. 14A Testis

a. Fig. 14A-Testis DAPI 100X_RGB_blue;

b. Fig. 14A-Testis Ab 100X_RGB_red;

c. Fig. 14A-Testis GFP 100X_RGB_green;

d. Fig. 14A-Testi overlap 100X_RGB.

17. Fig. 14B Fshr expression in mouse organs by dd.PCR: one Excel file

18. Negative control B6: 8 sub-folders, 4 TIFF in each sub-folders

1) Aorta:

a. Negative control-Aorta IgG Ab 100X_RGB_red;

b. Negative control-Aorta IgG DAPI 100X_RGB_blue;

c. Negative control-Aorta IgG GFP 100X_RGB_green;

d. Negative control-Aorta IgG overlap 100X_RGB.

2) BAT

a. Negative control-BAT IgG Ab 100X_RGB_red;

b. Negative control-BAT IgG DAPI 100X_RGB_blue;

c. Negative control-BAT IgG GFP 100X_RGB_green;

d. Negative control-BAT IgG overlap 100X_RGB.

3) Bone

a. Negative control-Bone IgG Ab 100X_RGB_red;

b. Negative control-Bone IgG DAPI 100X_RGB_blue;

c. Negative control-Bone IgG GFP 100X_RGB_green;

d. Negative control-Bone IgG overlap 100X_RGB.

4) Lung

a. Negative control-Lung IgG Ab 100X_RGB_red;

b. Negative control-Lung IgG DAPI 100X_RGB_blue;

c. Negative control-Lung IgG GFP 100X_RGB_green;

d. Negative control-Lung IgG overlap 100X_RGB.

5) Muscle

a. Negative control-Muscle IgG Ab 100X_RGB_red;

b. Negative control-Muscle IgG DAPI 100X_RGB_blue;

c. Negative control-Muscle IgG GFP 100X_RGB_green;

d. Negative control-Muscle IgG overlap 100X_RGB.

6) Ovary

a. Negative control-Ovary IgG Ab 100X_RGB_red;

b. Negative control-Ovary IgG DAPI 100X_RGB_blue;

c. Negative control-Ovary IgG GFP 100X_RGB_green;

d. Negative control-Ovary IgG overlap 100X_RGB.

7) Testis

a. Negative control-Testis IgG Ab 100X_RGB_red;

b. Negative control-Testis IgG DAPI 100X_RGB_blue;

c. Negative control-Testis IgG GFP 100X_RGB_green;

d. Negative control-Testis IgG overlap 100X_RGB.

8) Thyroid

a. Negative control-Thyroid IgG Ab 100X_RGB_red;

b. Negative control-Thyroid IgG DAPI 100X_RGB_blue;

c. Negative control-Thyroid IgG GFP 100X_RGB_green;

d. Negative control-Thyroid IgG overlap 100X_RGB.

Access information

Other publicly accessible locations of the data:

Data was derived from the following sources:

Fshr-ZsGreen GFP reporter mice
B6 mice.