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. 2021 Aug 31;297(4):101152. doi: 10.1016/j.jbc.2021.101152

Figure 2.

Figure 2

Effect of isoform-specific PKD siRNA transfection on tissue factor (TF) expression and TF surface activity induced by LPA in MASMCs.A, MASMCs were treated with 10 μM of LPA for various time points and TF expression was detected by Western blotting analysis with TF antibody. PKD1 protein expression in the time course was served as an internal control (PKD1 image is the same shown in Fig. 1B). B, effect of knockdown of the expression of PKD isoforms with PKD isoform-specific siRNAs on LPA-induced TF protein expression. Expression of PKDs and TF proteins was examined by Western blot analysis. For each PKD isoform, two specific siRNAs targeting different sequences were utilized. The specific PKD siRNAs (40 nM) were transfected into MASMCs for 48 h followed by serum starvation for 24 h; then cells were stimulated with 10 μM of LPA for 5 h. β-actin serves as a loading control. C, knockdown results of panel B were quantified by densitometry. Data are mean ± SD from three experiments. ∗∗p < 0.01 versus LPA-treated, nonsilencing siRNA group. D, effect of knockdown of the expression of PKD isoforms with PKD isoform-specific siRNAs on LPA-induced TF surface activity. Data are mean ± SD from three experiments. ∗∗p < 0.01 versus LPA-treated, nonsilencing siRNA group.