Figure 2

ZIP13^−/− cell lines reveal autophagy impairment. (a) Microscopic analysis reveals that after 3 days in culture without medium change, floating and spherical cells appear in the ZIP13^−/− cultures. (b) Western blot analysis reveals the downregulation of LC3 proteins in the ZIP13^−/− cell line. Uncropped blots are shown in Fig. S1b. (c) Confocal microscopic analysis reveals the decrease in LC3-positive vacuoles in the ZIP13^−/− cell line after 3-day culturing without medium change (n = 5, ***P < 0.005). Scale bars = 25 μm. (d) Western blot analysis shows that bafilomycin treatment does not restore the accumulation of LC3 proteins in the ZIP13^−/− cell line. The relative intensity of total LC3s was measured by the ImageJ software (http://rsbweb.nih.gov/ij/download.html). The data represent three independent experiments (***P < 0.005). Uncropped blots are shown in Fig. S2.