Fig. 1: Testicular atrophy in Kdm2a iKO animals.
A Schematic diagram illustrating the targeting and conditional deletion strategies of the Kdm2a locus. F1 and R1 represent PCR primers used for genotyping. Locations of FRT and loxP sites are indicated. B Tamoxifen dosing schedule of Ctrl (Kdm2a2lox/2lox) and Kdm2a inducible knockout (iKO: Kdm2a2lox/2lox; ActbCreERT2/+) animals. C Testes of Ctrl and Kdm2a iKO males at 65 d post first tamoxifen treatment. Scale bar = 2 mm. D Combined testes weights of Ctrl and iKO mice at 65 d post first tamoxifen treatment. n = 6 males per group. Two-sample, two-sided Wilcoxon tests were performed between indicated groups ***P ≤ 0.001. Center lines in box plots represent median values, while lower and upper lines indicate the interquartile range (IQR; from the 25th to 75th percentile). E Kdm2a genotyping results of total testes genomic DNA showing progressive loss of the Kdm2a2lox allele over time upon tamoxifen administration. (n = 1 animal per timepoint). However, at least three animals showed the maximal biological effect (complete excision of exon 8 at day 15). F Immunofluorescence staining of testicular sections from Ctrl (Kdm2a2lox/2lox) (n = 1) and Kdm2a inducible knockout (iKO: Kdm2a2lox/2lox; CAGG-Cre-ERTM) (n = 1) at 9 d post first tamoxifen treatment with anti-c-KIT (green) and anti-KDM2A (red) antibodies. Acrosomes and nuclei were stained with Lectin PNA (blue) and DAPI (white), respectively. Seminiferous tubule borders are highlighted with yellow dashed lines. Yellow asterisk in interstitial space marks c-KIT-positive Leydig cells. The yellow arrowhead points to a KDM2A-positive SgU lacking c-KIT expression. Scale bar = 50 μm. G Scheme adapted from2 and (74) summarizing expression and localization of KDM2A protein (red) in relation to developmental progression of male germ cells (related to Supplementary Fig. 2b). H PAS-stained testis section of Ctrl (stage VIII tubule) (n = 6) and Kdm2a iKO (n = 6) males, 65 d post first tamoxifen treatment. Scale bar = 50 μm.
