Figure 5
Erns acts as a receptor of mitophagy to interact with LC3-II. A HEK 293T cells were transfected with Flag-LC3-II and HA-ERNS. Cell lysates were immunoprecipitated with anti-Flag antibody and then analyzed by Western blot. B Western blot analysis was performed to detect VDAC1, MFN2, p62 and LC3-II expression in empty vector control group, NCP BVDV group and ERNS overexpression group at 48 hpi. C IFA analysis of the recruitment of ERNS to damaged mitochondria in MDBK cells stably expressing ERNS at 48 hpi. NCP BVDV-infected (MOI = 5) cells were used as positive control. (Scale bar = 10 μm). D The sequences of the LIR motif (W/YxxI/L) in ERNS were aligned manually with typical autophagy receptors. E HEK 293T cells were co-transfected with each HA-tagged WT ERNS and its LIR mutants (ERNS W84A+I87A and ERNSΔLIR) with Flag-LC3-II. Cell lysates were immunoprecipitated with anti-Flag antibody and then analyzed by Western blot. F HEK 293T cells were transfected with HA-WT ERNS and its LIR mutants. Cell lysates were harvested for Western blot analysis at 48 hpi. Data are given as means ± standard deviation (SD) from three independent experiments. P values were calculated using Student t test. An asterisk indicates a comparison with the indicated control. *P < 0.05; **P < 0.01; ns: not significant.