Fig. 1: Golgi stress induces proteasomal degradation of GM130.
From: Golgi organization is regulated by proteasomal degradation
a HeLa cells were treated with either cycloheximide (CHX 100 µg/ml, 8 h), CHX and MG-132 (20 µM, 8 h) or left untreated and analyzed by WB with the indicated antibodies. b–e HeLa cells expressing an inducible knockdown shRNA vector targeting PSMD6 were either treated with DOX to induce shRNA expression, or left untreated. b The expression levels of PSMD6, GM130, or K48-linked poly-ubiquitination were evaluated by WB analysis. Image is representative of three experiments. p(PSMD6) = 0.0001; p(K48) = 0.0011; p(GM130) = 0.0175. c–e Band intensities relative to actin were quantified using ImageLab. n = 3 independent experiments. Error bars = SD. p(PSMD6) = 0.0065, p(K48) < 0.0001,p(GM130) = 0.0039 (t-test, log relative intensity). f, g A549 cells stably expressing an inducible shRNA for PSMD6 were transfected with siRNA targeting wither SLC35A1, CMAS, or non-targeting control and incubated in the presence of either DOX (to induce PSMD6 shRNA) or DMSO as control for 72 h. f Representative images of Giantin immunostaining in the different cells. Scale bar = 50 μM. g The relative Golgi size was determined based on Giantin staining. Each dot represents a single cell. Error bars = SD. n = 1000 cells; *p = 0.0123; ****p < 0.0001 (one way ANOVA with Sidak’s multiple comparisons test). h Cells treated as in F were analyzed by WB for GM130 expression. Source data are provided as a Source Data file.
