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. Author manuscript; available in PMC: 2020 Jun 1.
Published in final edited form as: Arterioscler Thromb Vasc Biol. 2019 Jun;39(6):1212–1226. doi: 10.1161/ATVBAHA.119.312729

Figure 6. PLCβ3 mediate thrombin-induced IL-33 expression and HASMC migration.

Figure 6.

A & B. HASMCs were transfected with the indicated siRNA (100 nmoles), quiesced, treated with and without thrombin (0.5 U/ml) for 6 hrs and either RNA was isolated or cell extracts were prepared. The RNA and cell extracts were analyzed by RT-PCR and Western blotting, respectively, for IL-33 and β-actin mRNA and protein levels using their specific primers or antibodies, respectively. Cell extracts were also analyzed by Western blotting for PLCβ3 levels to show the efficacy of its siRNA. C. HASMCs were transfected with the indicated siRNA, quiesced and subjected to migration assay. D. Blockade of Par1 by its antagonist SCH79797 or depletion of Gαq/11 or PLCβ3 levels attenuate thrombin-induced NFATc1 translocation from cytoplasm to nucleus. E. Time course effect of thrombin on complex formation of NFATc1 with E2F1 and LMCD1. F. All the conditions were the same as in panel B except that siControl and siPyk2 were tested on thrombin-induced IL-33 levels and the blot was reprobed for Pyk2 and β-actin levels. The bar graphs represent Mean ± S.D. values of three independent experiments. *, p < 0.05 versus siControl; **, p < 0.05 versus siControl + thrombin.