Fig. 2
Effect of knockdown and transfection of AGR2 on cellular TXT sensitivity. Treatment of UT-SCC-70 cells with AGR2-specific siRNAs (HSS173724 and HSS16220) significantly repressed cellular AGR2 expression both at 24 and 72 hrs after initiation of the treatment. A. The repression of AGR2 expression was associated with a decrease of IC50 values of TXT (docetaxl), thereby sensitizing the cells to the drug (B). In contrast, AGR2 transfection into UMB-SCC-745 cells enhanced the expression both in the gene and protein expression levels (C), which increased IC50 values of TXT in the stable AGR2 transfectants (#1 and #6). The enhanced cellular TXT resistance was well associated with the enhanced AGR2 expression (D). Western blot analyses and quantification of the blots were performed for AGR2 and β-actin and the blots displayed were cropped from each transferred membrane (C) (See Supplemental Information File 1_Western Blot Raw Data). These expression-drug sensitivity associations were not always observed against 5-FU (5-fluorouracil), although AGR2 was suggested to be involved in also 5-FU sensitivity (E): AGR2 expression was normalized with the geometric mean of GAPDH, ACTB and HPRT1 expression and showed as relative expression to that in Negative Control (NC); Error bars represent standard errors (SE); IC50, inhibitory drug concentration of 50% cell growth or drug concentration of 50% optical density of control; Relative IC50 value of TXT (%) represents the ratio to that in control sample