Figure 3.

Non-elicited peritoneal and BM-derived atg7^−/- macrophages do not conjugate PE to LC3. (A) Comparison of LC3 protein expression in WT, atg7^−/-, and atg5^−/- peritoneal macrophages elicited by thioglycolate (thio), proteose-peptone (pp), and concanavalin A (cc). (Ai) Representative LC3 immunoblot with ACTB/β-actin as a loading control. (Aii) Mean LC3 expression values (+ SD) obtained from four individual mice. One-way ANOVA with Bonferroni correction were used to calculate differences between the different treatments; * p < 0.05, ** p < 0.01, *** p < 0.001. Student’s t-test (two-tailed) with Welch correction was used to compare LC3-II with LC3-I; ^# p < 0.05, ^## p < 0.01. (B) Representative LC3 immunoblotting in non-elicited peritoneal (P-), thioglycolate (thio)-elicited (P+), and BM-derived (BM-) WT, atg5^−/-, and atg7^−/- macrophages, respectively. Bars represent mean LC3-II:LC3-I ratios + SD (n = 5). Statistics were calculated using one-way ANOVA with Bonferroni correction. * differences by treatment; ^# differences by genotype; * p < 0.05, **^/## p < 0.01, *** p < 0.001. LC3 immunoblotting in (Ci) BM- and (Cii) P- WT and atg7^−/- macrophages treated with lipopolysaccharide (LPS), IFNG (interferon gamma), thioglycolate (thio), conditioned media (cm) collected after incubation of thioglycolate-elicited WT macrophages for 24 h, or left untreated (C). ACTB/β-actin was used as loading control