Phenylhydrazonopyrazolone sulfonate 1 phps1

Phenylhydrazonopyrazolone sulfonate 1 (PHPS1) was obtained from Sigma-Aldrich (St. Louis, MO). SB-3CT was obtained from Selleckchem (Houston, TX). RPMI 1640, FBS, penicillin, and streptomycin were obtained from HyClone (Logan, UT). TRIzol reagents were purchased from Takara Bio, Dalian, China. All PCR primers were purchased from Shanghai Bioengineering (Shanghai, China). ERK, p-ERK, Shp2, p-Shp2, Smad2, p-Smad2, Smad3, p-Smad3, JNK, P-JNK, P38, p-P38, MMP-9, MMP-2, E-cadherin, vimentin, α-SMA and β-actin primary antibody were from Cell Signaling Technology (Danvers, MA), and Shp2 primary antibody was from Santa Cruz Biotechnology (Santa Cruz, CA). MMP-9 and TGF-β1 ELISA kits were from Boster (Wuhang, China). Lipofectamine LTX was from Invitrogen (Carlsbad, CA).

Recombinant human TGFβ1 (mammalian) was from PeproTech (Rocky Hill, NJ). Phenylhydrazonopyrazolone sulfonate 1 (PHPS1) was from Sigma-Aldrich. HOOK1 siRNA (target sequence: GUUGAGAUAUAUCGUCAGA) was from Thermo Fisher Scientific (Dharmacon Products, Lafayette, CO). SHP2 siRNA and all siRNA negative controls were from Santa Cruz Biotechnology Inc. (Santa Cruz, CA). Antibodies for E-cadherin, vimentin, SHP2 and Hook1 were from Santa Cruz Biotechnology. Antibodies for pERK, ERK, pSmad2 (Ser467), pSmad3 (Ser423/425), Smad2, Smad3, COL1A1, and Snail1 were from Cell Signaling Technologies (Beverly, MA). Anti-Myc monoclonal antibody was from OriGene (Rockville, MD). Anti-Flag® antibody produced in rabbit was from Sigma-Aldrich. IRDye 680LT/IRDye 800CW secondary antibodies were from LI-COR Biosciences (Lincoln, NE). Primer synthesis and DNA sequencing were performed by Invitrogen.