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. 2019 Dec 6;11(12):1959. doi: 10.3390/cancers11121959

Figure 3.

Figure 3

Effect of SMAD3 knockdown on basal migratory activity in Panc1 cells. (A) Panc1 cells were transiently transfected twice with 50 nM of either control (Co) siRNA or SMAD3 siRNA. Forty-eight h after the second round of transfection, cells were subjected to real-time cell migration assay. Inset, functional validation of the SMAD3 siRNA by immunoblotting. siCo, Co siRNA; siS3, SMAD3 siRNA. (B) Panc1 cells were transiently transfected twice with 25 nM each of Co siRNA for SMAD3 + Co siRNA for RAC1B, RAC1B siRNA + Co siRNA for SMAD3, or RAC1B siRNA + SMAD3 siRNA. Forty-eight h after the second round of transfection cells were subjected to real-time cell migration assay as in (A). Inset, Quantitative comparison of SMAD3 and RAC1B knockdown on migratory activities. Data shown are the mean ± SD (n = 4). Differences in the mean values among the three treatments did not reach statistical significance. Data in (A,B) are from one representative experiment out of four experiments with very similar results, and are the mean ± SD from 3–4 wells per condition. In (A), differences between Panc1 + SMAD3 siRNA (red curve, tracing B) and Panc1 + Co siRNA (blue curve, tracing A) are significant at 0:30 and all later time points (p < 0.05, Mann-Whitney U-test). In (B), differences between Panc1 + RAC1B siRNA + SMAD3 siRNA (red curve, tracing C) and Panc1 + Co siRNAs (blue curve, tracing A) are significant at 5:30 and all later time points (p < 0.05, Mann-Whitney U-test).