Anti sncg

TGF-β was purchased from PeproTech (USA). The antibodies used were as follows: anti-Twist1 was purchased from Abcam (Cambridge, UK); anti-SNCG and anti-β-actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA); anti-Smad2 and anti-Smad3 were purchased from ABclonal (Wuhan, China); anti-phospho-Smad2 (S245/250/255) and anti-phospho-Smad3 (S423/425) were from Cell Signaling Technology (Danvers, MA, USA). All siRNAs were custom-synthesized products of Ribobio Co., Ltd. (Guangzhou, China). The siR-Ribo negative control (siControl) was used for all siRNA experiments. The target sequences for Twist1 knockdown are as follows: 5′-GGUACAUCGACUUCCUCUA-3′ for siTwist1#1, and 5′-UUGAGGGUCUGAAUCUUGCUCAGCU-3′ for siTwist1#2. The sequence of siRNA targeting the 3′-untranslated region of Twist1 gene (siTwist1#3) is 5′-CACCTCTGCATTCTGATAGAA-3′. The two target sequences for SNCG knockdown are as follows: 5′-GCAGCTGAGAAGACCAAGG-3′ for siSNCG#1 and 5′-GGAGAATGTTGTACAGAGC-3′ for siSNCG#2. Target sequences for Smad3 are 5′-GGAGAAAUGGUGCGAGAAG-3′.

TGF-β was purchased from PeproTech (Rocky Hill, NJ, USA). The p38MAPK inhibitor SB203580 and MMP inhibitor BB-94 were from MedChemExpress (NJ, USA). The proteasome inhibitor MG132 was from Merk-Millipore (Darmstadt, Germany). Cycloheximide was from Beyotime Biotechnology (Shanghai, China). The antibodies used were as follows: anti-Twist1, anti-FLAG, anti-phosphorylated MKK3 (S189) and anti-phosphorylated MKK6 (S207) (Abcam, Cambridge, UK); anti-SNCG and anti-β-actin (Santa Cruz Biotechnology, Santa Cruz, CA, USA); anti-Erk1/2, anti-phosphorylated Erk1/2 (T202/Y204) and anti-phosphorylated p38MAPK (T180/Y182) (Cell Signaling Technology, Beverly, MA, USA); anti-p38α and anti-vimentin (Proteintech, Rosemont, IL, USA); anti-MMP-9, anti-SMAD3, anti-MKK3/6, anti-fibronection, anti-N-cadherin (Abways Technology, Shanghai, China). The pcDNA3-Twist1 and PCI-SNCG plasmids were prepared as described previously 12 (link), 20 (link).