Fig. 6
A peroxynitrite decomposition catalyst rescues the combination effects on ER stress, cytoplasmic vacuolization and viability. (A, Ai) Western Blot analysis of BiP/GRP78 in NCH82 cells treated with the vehicle control (DMSO) and 48 µM TMZ in combination with 2.66 µM SKI-II (TMZ + SKI-II) in absence or presence of Bafilomycin A (BA1), for 24- to 48 h under normoxia (21% O2) and hypoxia (3% O2). GAPDH was used as a loading control. A Representative blot of cells treated under 3% O2. Full-length blots are presented in Additional file 9. (Ai) BiP protein expression relative to GAPDH was quantified with ImageJ. B Gene expression analysis of DDIT3 in NCH82 cells treated accordingly for 24 h under normoxia and hypoxia. Data represent mean (± SD) of n = 3. Two-way ANOVA followed by Tukey’s multiple comparisons test was performed for statistical analysis. C Western blot analysis of BiP/GRP78 in NCH82 cells treated accordingly for 24 h under normoxia (21% O2) and hypoxia (3% O2). FeTPPS was used at a final concentration of 100 µM. A representative blot is shown. Full-length blots are presented in Additional file 10. (Ci) BiP/GRP78 protein expression level was quantified and normalized to GAPDH with ImageJ. D Viability of cells treated accordingly for 72 h was determined via the PrestoBlueTM assay. Data were normalized to the DMSO condition. Data represent mean (± SD) of n = 3. Two-way ANOVA followed by Tukey’s multiple comparisons test was performed for statistical analysis