Fig. 5

β-catenin regulated the expression of SPC markers and HDACs. The mRNA expression of Hdac1-9 was determined in SPCs using RT-PCR (T: testis; S: SPCs; N: negative control) (A). Expression of HDAC4 in testis was detected from 42-day old mouse using IHC (B). Co-IF staining demonstrated the expression and colocalization of PLZF and HDAC4 in purified SPCs (C–G). The binding of β-catenin to HDAC4 and STAT3 was examined using Co-IP (H). The expression levels of β-catenin, AXIN2, Cyclin D1, HDAC4, GFRA1, PLZF, c-Kit, PCNA, BAX, BCL-2 and β-tubulin were determined in SPCs treated with scrambled or β-catenin siRNA using Western blot, n=3 (I), and were statistically analyzed (J). Western blot was employed to detect the expression levels of HDAC4, PLZF, GFRA1, PCNA, BAX, BCL-2, AXIN2, c-Kit, STRA8, SOHLH2 and β-tubulin in scrambled or Hdac4 siRNA transfected SPCs, n=3 (K), and the results were statistically analyzed (L). The expression levels of STAT3, PLZF, STRA8, PCNA, BAX, BCL-2, AXIN2 and β-tubulin were determined in SPCs transfected with scrambled or Stat3 siRNA, n=3 (M), and were statistically analyzed (N). Scale bar = 20 μm, data represent as mean ± SD, *p<0.05,**p<0.01