Fig. 3.

Quercetin promoted GEM-induced cytotoxicity by suppressing RAGE expression in MIA Paca-2 cells. Effect of quercetin on (A) cytotoxicity, (B) RAGE mRNA expression, and (C) RAGE protein levels in Mia Paca-2 cells. (D) Cells were pretreated with various concentrations (6.25, 12.5, 25 and 50 μM) of quercetin for 24 h and then treated with 500 nM GEM for 72 h. Cell viability was measured via the MTT assay. (E) Autophagy-associated vacuolation in cells. (F) Percentages of autophagosomes. (G) Autophagy-associated ATG5, Beclin-1, LC3I, and LC3II protein expression. (H) Apoptotic cells were stained with Annexin V-FITC positive/PI-negative staining. (I) Caspase 3 activity. (J) Protein levels of p65. (K) Effect of quercetin on mean calcein fluorescence intensity in MIA Paca-2 cells. Reported values are means ± SD (n = 3), *p < 0.05 versus the control. MFI: mean fluorescence intensity.