Fig 1: MiR-483 promotes TGF-ß1-induced EMT and Tiam1/Rac signaling by downregulating Pard3. a Pard3 mRNA expression in the ATC cell lines, 8505C, HTH7, and FRO, and normal thy-ori 3-1 cells (*p < 0.05, **p < 0.01, one-way analysis of variance). b Knockdown of Pard3 (Pard3-shRNA1–3) in FRO cells and qRT-PCR assessment of Pard3 expression (*p < 0.05, **p < 0.01, ***p < 0.001, one-way analysis of variance). c–e FRO cells were stably transfected with Pard3-shRNA3 or scr. E-cadherin, vimentin, Tiam1, and Rac1 expression were detected by western blotting. GAPDH was used as a loading control (**p < 0.01, Student’s t test). The 8505C cells were stably transfected with pcDNA3.1-Pard3 and subsequently treated with TGF-ß1 (10 ng/mL) for 48 h. Untransfected cells with or without TGF-ß1 treatment were also included. f–h E-cadherin, vimentin, Tiam1, and Rac1 expression were detected by western blotting. GAPDH was used as a loading control (*p < 0.05, **p < 0.01, one-way analysis of variance). FRO cells were transfected with Pard3-shRNA3, and subsequently treated with TGF-ß1 (10 ng/mL) for 48 h. Untransfected cells with or without TGF-ß1 treatment were also included. i–k E-cadherin, vimentin, Tiam1, and Rac1 expression were detected by western blotting. GAPDH was used as a loading control (*p < 0.05, **p < 0.01, one-way analysis of variance). ATC cells stably transfected with pcDNA3.1-NC or pcDNA3.1-Pard3 were transfected with miR-483 inhibitor/miR-483 inhibitor NC, or miR-483 mimics/miR-483 mimics NC, and subsequently treated with TGF-ß1 (10 ng/mL) for 48 h. Untransfected cells with or without TGF-ß1 treatment were also included. E-cadherin, Tiam1, and Rac1 expression were detected by western blotting in both 8505C cells (l–n) and FRO cells (o–q). GAPDH was used as a loading control (**p < 0.01, ***p < 0.001, one-way analysis of variance, NS: non-significant). N = 3 independent experiments with triplicate biological replicates for each line
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