Fig. 4. BiP contributed to cell death induced by Rom+Cis+Gem.
a J82 and J82-Ras cells were treated with Rom+Cis+Gem at their IC10 doses for 0, 6, 12, 24, and 48 h. b J82 cells were constantly transfected with a BiP expression vector to result in J82-BiP-1 and -2 cell lines. J82 cells were constantly transfected with BiP-specific shRNA vectors to result in J82-shBiP-1 and -2 cell lines. c, d J82-BiP-1, BiP-2, shBiP-1, and shBiP-2, as well as J82-Ras cells were treated Rom+Cis+Gem. a–c Cell lysates were prepared and analysed by immunoblotting using specific antibodies to detect the levels of BiP with β-actin as a control, and these levels were quantified by densitometry. Levels of BiP (BiP/actin) were calculated by normalising with the level of β-actin, and the level set in control cells as 1 (X, arbitrary unit). d Cell viability was determined, and relative cell viability was normalised by the value determined in untreated counterpart cells, set as 100%. Columns, mean of triplicates; bars, SD. p Value was adjusted for multiple comparisons by using the Simes method. Statistical significance is indicated by **p < 0.01, ***p < 0.001. All results are representative of three independent experiments.