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. 2022 Aug 25;11(17):2645. doi: 10.3390/cells11172645

Figure 6.

Figure 6

SIRT4 can modulate autophagic function in vivo and in vitro. (A) Genotyping identification of SIRT4-knockout mice. (B,C) Autophagic flux in retinas of wild-type and SIRT4-knockout mice was monitored by LC3-II/I conversion with anti-LC3 and SQSTM1/p62 antibodies. (D) The overexpression efficiency of the SIRT4 plasmid was validated by western blotting. (E,F) Autophagic flux in P-CON and P-SIRT4 r-MCs was monitored by LC3-II/I conversion with anti-LC3 and SQSTM1/p62 antibodies. Data are shown as means ± SEMs. (G) The knockout efficiency of the SIRT4 lentivirus was validated by western blotting. (H,I) Autophagic flux in V-CON and V-SIRT4 r-MCs was monitored by LC3-II/I conversion with anti-LC3 and SQSTM1/p62 antibodies. Data are shown as means ± SEMs. (J) Autophagic puncta in MGCs were obtained by immunostaining with an LC3 antibody (scale bar: 20 μm). (K) Quantitative analysis of autophagic puncta in MGCs. Data are shown as means ± SEMs. (n = 3 per group; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001). SIRT4, sirtuin 4; WT, wild type; KO, knockout; p62, SQSTM1/p62.