Research Paper Volume 10, Issue 11 pp 3104—3116

Inactivation of glycogen synthase kinase-3α is required for mitochondria-mediated apoptotic germ cell phagocytosis in Sertoli cells

Figure 4. GSK3α participates in Sertoli cell phagocytosis of apoptotic germ cells. (A-B) Western blots and histogram showing the protein levels of GSK3α and p-GSK3α in control and HS treated TM4 cells. C: control; HS: heat shock. (C) The phagocytosis of Indian ink beads by TM4 cells observed by light microscopy. Black dots indicate engulfed Indian ink in TM4 cells. (D) Quantification of Indian ink beads phagocytosis via Micro plate spectrophotometer read as an OD value. (E) Histogram showing percentage of TM4 cells engulfing apoptotic germ cells as derived from immunofluorescence analysis. (F) Immunofluorescence analysis showing phagocytosis of apoptotic germ cells by TM4 cells treated with HS or GSK3α inhibitor. TM4 cells were fed with apoptotic male germ cells labeled with DAPI. Con: control, HS: heat shock, SB: SB216763. Thick arrow indicates engulfed germ cells. Thin arrow indicates unengulfed germ cells. Scale bars=10 μm. Values are expressed as the mean±SEM, n=15. Values with different superscripts are significantly different from each other (P<0.05).