Fig. 3: Spata33 knockout inhibits autophagy.
From: SPATA33 is an autophagy mediator for cargo selectivity in germline mitophagy
a LC3B puncta were detected in WT TM4 cells after starvation culture (EBSS) for 0.5 h. The cells were cultured in EBSS for the indicated time and analyzed by immunofluorescence with the anti-LC3B antibody and confocal microscopy. The insets showed an enlarged view of the indicated squares and highlighted red LC3B puncta in the cytoplasm. The nuclei were stained with Hoechst reagent. Scale bar: 20 μm. b LC3B puncta were detected at 1 and 2 h under starvation culture in the Spata33−/− cells. Scale bar: 20 μm. c To rescue Spata33 expression in the Spata33−/− cells, the cells were infected with lentivirus expressing 3xFLAG-SPATA33. Immunofluorescence with the anti-LC3B antibody showed obvious LC3B puncta appeared again at 0.5 h under starvation culture. Scale bar: 20 μm. d To overexpress Spata33 in the WT TM4 cells, the cells were infected with lentivirus expressing 3xFLAG-SPATA33 and analyzed by immunofluorescence with the anti-LC3B antibody. Obvious LC3B puncta were detected again at 0.5 h under starvation culture. Scale bar: 20 μm. e. Statistical analysis of LC3B puncta from (a) to (d). The number of LC3B puncta in 30 cells was counted for each group. Data are presented as mean ± S.D. *p < 0.05; **p < 0.01 (n = 3 independent experiments).
