Fig. 2.

Transgene expression and DAZL knockdown. (A) Western blot analysis to detect DAZL-MYC production in cells transduced with virus carrying pLLU2G (Vector) or pLLU2G-Dazl1 (Dazl-shRNA) and transiently transfected with pCDNA-Dazl-myc. The blot was stripped and reprobed with anti-tubulin to verify equal loading of protein. (B–E) Fluorescence of wild-type (B and D) and transgenic (C and E) neonatal pups (B and C) or adult ear punch skin (D and E). (F) RNase protection assay using a probe to detect Dazl short interfering RNA (from Dazl shRNA 1) in testis RNA from transgenic rats (17-9 or 16-13 Tg), a wild-type sibling of the 17-9 transgenic rat (Wt), or yeast RNA (mock). The protected portion of the antisense probe sequence is shown in red in Fig. 1. The sense probe is reverse-complementary to the antisense probe. Addition of RNase is indicated by +. (G) Western blot analysis to detect Dazl, Tex11, tubulin, and GFP expression in the testis. Lanes were loaded as follows. (Left) First lane, transgenic 16-13; second lane, wild-type sibling of 17-9 F₁; third and fourth lanes, two transgenic 17-9 F₁s. (Right) First lane, wild-type sibling of 17-9 F₂; second lane, transgenic 17-9 F₂. (H–K) Immunofluorescence to detect DAZL protein in testis cryosections of a Dazl-shRNA rat (H) and a wild-type sibling (J). DAPI (DNA) staining of the same sections is also shown (I and K).