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Fig. 7 | Cancer Cell International

Fig. 7

From: A drug screening assay on cancer cells chronically adapted to acidosis

Fig. 7

Autophagy and proteasomal activity are not affected by VP. a HCT116 and AA-HCT116 cells were exposed to VP for 4 and 8 h, or to EBSS, in presence or absence of BafA1 to analyse the turnover of LC3-II and SQSTM1 by WB. b Clustering analysis of the 229 genes associated with the four ubiquitinrelated GO terms found to be significantly enriched in the GO analysis. c HCT116 and AA-HCT116 cells were exposed to VP or BZ for 4 h and the presence of poly-Ub conjugates and free Ub was detected by WB. d, e HCT116 and AAHCT116 cells were treated with VP or BZ for 4 h and cell extracts exposed to the fluorogenic substrate Suc-LLVY-AMC to measure chemotrypsin-like activity of the proteasome. f The cell line MelJuSo UbG76 V-YFP was used as reporter of proteosomal inhibition after treatment with VP or BZ. Accumulation of Ub-YFP indicates inhibition of proteasomal activity. g The levels of CDKN1A were analysed as marker of proteasomal activity in HCT116 and AA-HCT116 cells treated with BZ or VP at the indicated times and concentrations

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