Figure 4
Basal autophagic degradation of MyD88 is mediated by a mechanism different from macroautophagy. (a) Treatment with the inhibitors of macroautophagy does not affect basal MyD88 expression. MEFs were incubated with 0.5% DMSO (control), 10 mM 3-MA, 100 nM wortmannin, or 10 μM spautin-1 for 12 h. Expression levels of MyD88 and α-tubulin were assessed by immunoblotting. All the blots were obtained under the same experimental conditions, and the cropped images of the blots are shown. The uncropped images are in Supplementary Fig. 20. (b) Deficiency in ATG5 does not affect BafA1 treatment-increased basal MyD88 expression. Atg5+/+ MEFs and Atg5−/− MEFs were incubated with 0.5% DMSO (BafA1−) or 100 nM BafA1 (BafA1+) for 12 h. Expression levels of MyD88 and α-tubulin were assessed by immunoblotting. All the blots were obtained under the same experimental conditions, and the cropped images of the blots are shown. (c) Starvation-induced degradation of basal MyD88 is suppressed by the inhibitors of macroautophagy. MEFs were incubated with or without 10 mM 3-MA or 10 μM spautin-1 for 6 h, and then cultured under the starvation conditions for the indicated periods. Expression levels of MyD88 and α-tubulin were assessed by immunoblotting. All the blots were obtained under the same experimental conditions, and the cropped images of the blots are shown. (d) Starvation-induced degradation of basal MyD88 is affected by ATG5 deficiency. Atg5+/+ MEFs and Atg5−/− MEFs were cultured under the starvation conditions for the indicated periods. Expression levels of MyD88, LC3, ATG5, and α-tubulin were assessed by immunoblotting. Values within parentheses represent the ratio determined by densitometric measurement of the bands. All the blots were obtained under the same experimental conditions, and the cropped images of the blots are shown. The uncropped images are in Supplementary Fig. 21.
