Fig. 4

Pharmacological inhibition of RPA disrupts metabolic activities. a S26 and 5-8F cells were subjected to 24- or 48-h treatment with DMSO or 10 μM HAMNO, followed by immunoblot analysis to assess pmTOR, pBeclin1, total Beclin1, LC3B, ATG5, p62, cleaved caspase 3 and ACTIN levels. b S26 and 5-8F cells were treated with increasing doses of HAMNO (concentrations in μM shown) for 24 h. Relative gene expression of WIP1, SQSTM1 and GABARAPL1 was analyzed by qRT-qPCR. mRNA levels were normalized to ACTB mRNA. Relative expression quantified via the double delta Ct method is plotted. Data are representative of three biological replicates. c Effects of RPAi treatment at the indicated dose for 24 h on glucose utilization, lactic acid production, and intracellular ATP levels in 5-8F and S26 cells. Data are representative of five biological replicates. d S26 and 5-8F cells were treated with HAMNO or cultured in medium containing dialyzed FBS and no glucose (16 h). Quantification of LC3B staining is presented for three biological replicates