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. 2012 Jul 13;41(4):1464–1474. doi: 10.3892/ijo.2012.1553

Figure 2.

Figure 2

Reduction of APLP2 and/or APP by siRNA impairs the growth of a pancreatic cancer cell line. S2-013 cells were transfected with pooled siRNA against APP and APLP2, alone or in combination. Non-targeting pooled siRNA was used as the negative control (control siRNA). (A) Western blot analysis confirmed specific knockdown of glycosaminoglycan (GAG)-modified APLP2, full-length APLP2, APLP2 C-terminal fragments and/or full-length APP (mature, top band; immature, bottom band) actin 48 h post-transfection. Actin was used as a loading control. APP siRNA does not affect APLP2 expression, and vice versa. (B) S2-013 cells were seeded 48 h post-transfection at low density and cell growth was determined by the MTT assay. MTT-specific absorbance (y-axis) was obtained by subtracting the absorbance at 690 nm from the absorbance at 570 nm; time is displayed as hours post-seeding. (C) Reduction of APP and APLP2 significantly reduced the growth of S2-013 compared to cells treated with control siRNA. Percent inhibition in growth was obtained from the data in (B) by normalizing the MTT-specific absorbance to control siRNA absorbance and seeding at 0 h. Asterisks in (B and C) denote significance by the analysis of variance (ANOVA), p<0.001; n=12, error bars indicate standard error of the mean.