LUBAC Formation Is Impaired in the Livers of Mice with MCD-Dependent Nonalcoholic Steatohepatitis

Figure 5

(a) HepG2 cells were incubated in the presence or absence of palmitate. After incubation for 12 hr, total RNA was extracted and reverse transcribed, and the mRNA levels of SHARPIN, HOIP, and HOIL-1L were quantified by real-time PCR. As the housekeeping gene, GAPDH was used for quantity normalization. The results shown are means ± S.E.M. (). n.s.: not significant. (b) HepG2 cells were incubated with the indicated combination of 20 μM MG132, 10 μM chloroquine, and 10 μM cycloheximide, in the presence or absence of palmitate for 24 hr. Cell lysates were harvested in lysis buffer and expressions of SHARPIN, HOIP, and HOI-1L were determined by immunoblotting using the corresponding antibodies. For each combination, a representative immunoblot is shown and quantified data are expressed as the percentage ± S.E.M of five independent experiments. (). n.s.: not significant.

(a)

(b)