Cytoplasmic and nucleus protein extraction kit

Total protein extraction: Cells were harvested by cytology brush, and lysed with RIPA lysis buffer (Sigma, USA) supplemented with phosphorylase and protease inhibitor mixture (Thermo, USA), quantified by the BCA assay.
Cytoplasmic and nucleus protein extraction: Cells were harvested by Tyrisin (Invitrogen), then cytoplasmic and nucleus protein was extracted by Cytoplasmic and Nucleus Protein Extraction Kit (Thermal Scientific, USA) according to its protocol, quantified by the BCA assay.
The standard detailed experimental process of the western blot was the same as our previous study (PMID29331423 and PMID29800682).

Cells were harvested by Tyrisin (Invitrogen), then cytoplasmic and nucleus protein was extracted by Cytoplasmic and Nucleus Protein Extraction Kit (Thermal Scientific, USA) according to its protocol, quantified by the BCA assay.
The standard detail experimental process of western blot as same as our previous study (PMID29331423 and PMID29800682). Western blot band was quantified through the Image-J software (NIH, USA). Antibodies against GAPDH, FHL3, E-cadherin, MMP2 and GSK3β were purchased from Proteintech (1:1000, Hangzhou, China), antibodies against smad2/3, smad4, twist1, Ubiquitin, Histone 3, ser473-phospho-Akt, try216/279-phospho-GSK3β, ser9-phospho-GSK3β and ser423/425-phospho-smad2/3 were purchased from Huabio (1:1000, Hangzhou, China), antibodies against ZO-1, Akt, vimentin, snail1 and zeb1 were gained from Abcam (1:1000, China).