DOI: 10.5061/dryad.1jwstqk4t
title: Atlas of FSHR expression from novel reporter mice

description: 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.
description: This dataset includes raw data collected by images of confocal
 microscope by IF and single RNA molecule-fluorescence in situ
 hybridization and dd.rtPPCR, etc., which were described in more details in
 Methods of our publication.
description: # Atlas of FSHR expression from novel reporter mice
 [https://doi.org/10.5061/dryad.1jwstqk4t](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: * NO Data was derived from the following sources: * Fshr-ZsGreen GFP reporter mice * B6 mice.