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. 2018 Oct 25;316(1):L216–L228. doi: 10.1152/ajplung.00538.2017

Fig. 6.

Fig. 6.

Overexpression of p53 downregulates Transient receptor potential channel (TRPC) attenuates SOCE and inhibits pulmonary arterial smooth muscle cell (PASMC) proliferation and migration, while downregulation of p53 upregulates TRPC channels, enhances SOCE, and increases PASMC proliferation and migration. A: Western blot analysis of p53, TRPC1, and TRPC6 in human PASMC transfected with an empty vector and p53. B: representative record showing changes in cytosolic Ca2+ concentration ([Ca2+]cyt) before, during, and after application of 10 µM cyclopiazonic acid (CPA) in the absence (0Ca) or presence of extracellular Ca2+ in human PASMC transfected with an empty vector and p53. The bar graph shows the amplitude of [Ca2+]cyt increases due to CPA-induced SOCE (means ± SE; n = 5). C: cell proliferation [bromodeoxyuridine (BrdU) incorporation; means ± SE; n = 5] and migration (%cells migrated; means ± SE; n = 5) in PASMCs transfected with vector and p53. D: Western blot analyses of p53, TRPC1, and TRPC6 in human PASMCs transfected with siRNA-NT and siRNA-p53. E: representative record showing changes in [Ca2+]cyt before, during and after application of CPA in the absence (0Ca) or presence of extracellular Ca2+ in human PASMCs transfected with siRNA-NT and siRNA-p53. The bar graph shows the amplitude of [Ca2+]cyt increases due to CPA-induced SOCE (means ± SE; n = 5). F: cell proliferation (BrdU incorporation; means ± SE, n = 5) and migration (% cells migrated; means ± SE; n = 5) in human PASMCs transfected with siRNA-NT and siRNA-p53. G: Western blot analysis of p53, STIM1, and STIM2 in human PASMC transfected with siRNA-NT and siRNA-p53 (left). Summarized data (means ± SE; right) showing the protein levels of p53, STIM1, and STIM2 in control PASMCs and siRNA-p53 treated PASMCs. H: Western blot analysis of p53, Orai1, and Orai2 in human PASMC transfected with siRNA-NT and siRNA-p53 (left). Summarized data (means ± SE; n = 5; right) showing the protein levels of p53, Orai1, and Orai2 in control PASMCs and siRNA-p53-treated PASMCs. For these studies, all cells used were between 5 and 8 passages. We compared the same passage number of cells for each experiment. *P < 0.05 and ***P < 0.001 vs. vector or siRNA-NT.