Fig. 5.
Cellular and molecular effects of 17-aminogeldanamycin (AG) activity used either alone or in combination with trametinib (TRA) in NRASQ61R melanoma cells a Changes in a viable cell number were assessed by acid phosphatase assay. Representative results are shown. *p ≤ 0.05 versus control b HSP70 and GRP78 transcript levels were assessed by qRT-PCR in cells incubated with 0.4 μM AG for 6 and 22 h, and expressed relatively to the control. *p ≤ 0.05 c Level of phospho-IRE1α (p-IRE1α) was determined after 4 and 24 h. β-actin was used as a loading control. Quantifications of p-IRE1α and IRE1α levels are shown below the blots. XBP1s transcript level was assessed by qRT-PCR after 22 h, and expressed relatively to control. d–g DMBC22 cells were incubated with 0.4 μM AG and 50 nM TRA used either alone or in combination. d Level of phosphorylated ERK1/2 (p-ERK1/2) was assessed by Western blotting, and β-actin was used as a loading control. e Percentages of Annexin V-positive cells after 24 and 48 h were determined by flow cytometry. Representative contour plots are shown. *p ≤ 0.05 drug combination vs. either drug used alone f Cleaved PARP (cPARP) was immunoblotted after 24 h. An equal loading was confirmed by β-actin. Quantification of cPARP level is shown below the blots. g Percentages of cells with active caspase-3/7 were assessed by time-lapse microscopy