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. 2016 Sep 1;12(9):e1006293. doi: 10.1371/journal.pgen.1006293

Fig 5. Mutant SSCs are lost due to defective maintenance.

Fig 5

(A-C) Controls treated with BrdU for 4 hours. IF of sectioned testes showing BrdU (red) and spermatogonial markers: Id4-GFP, PLZF or c-KIT (green). DAPI DNA stain (blue). Double-positive cells are indicated by the white arrowhead. Higher magnifications are shown in insets. (D) Quantification of BrdU and GFP+/PLZF+; GFP-/PLZF+ and c-KIT+ positive cells. Values are average from >300 tubule sections for GFP staining, and >100 for PLZF and c-KIT staining. Error bars indicate standard deviation. ** indicates P < 0.005 (Student’s T test). (E-G) Diagram of the BrdU chase experiment in the control (E) and two possible outcomes in the mutant: failure to maintain SSCs (F) and SSC death (G). (H, I) IF of sectioned testes showing tdTomato (red) and BrdU (green) in control (H) and mutant (I) 18 DPT with BrdU chase for 12 days. DAPI DNA stain (blue). Double-positive cells are indicated by the white arrowheads and tdTomato single-positive cells are indicated by yellow arrowheads. Scale bars: 20 μm.