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. 2018 Dec 26;294(8):2880–2891. doi: 10.1074/jbc.RA118.003822

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© 2019 Tian et al.

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Figure 7. — HS-152 does not interfere with TGFβ/SMAD pathway. A, HS-152 does not inhibit TGFβ-induced phosphorylation of SMAD2. MDCK cells pretreated 12 h with DMSO or 0.05 μm HS-152 were treated 20 h with or without 100 pm TGFβ and then subjected to immunoblotting of total cell lysates with indicated antibodies. B, HS-152 does not hinder the nuclear accumulation of SMAD2 in response to TGFβ treatment. MDCK cells pretreated 12 h with or without 0.05 μm HS-152 were treated another 20 h with or without 100 pm TGFβ, and then subjected to immunofluorescence assay. ZO-1 staining was detected with an anti-ZO-1 antibody (green), and SMAD2 was detected with an anti-SMAD2 antibody (red). C, HS-152 does not block TGFβ-induced vimentin expression. MDCK cells treated as in (B) were subjected to immunofluorescence assay to detect ZO-1 (green) and vimentin (red) staining using anti–ZO-1 and anti-vimentin antibodies, respectively. D, HS-152 blocks TGFβ-induced down-regulation of ZO-1 but not up-regulation of vimentin. MDCK cells treated as in (B) were subjected to immunoblotting assay with indicated antibodies.