Goat anti mouse igg polyclonal antibody

Using a Cell Mitochondria Isolation Kit (Beyotime, Shanghai, China), we extracted proteins from target cells. The samples were incubated with the following primary antibodies: anti-TGFβ1 (ab21610, Abcam, Cambridge, MA, USA), anti-TGFβ1 (ab64715, Abcam), anti-α-SMA (ab5694, Abcam), anti-p-SMAD2 (ab53100, Abcam), anti-SMAD2 (ab40855, Abcam), anti-MMP1 (ab52631, Abcam), anti-MMP3 (ab52915, Abcam), anti-ACTG2 (SAB1411364, Sigma-Aldrich, St. Louis, MO, USA), and anti-GAPDH (ab8245, Abcam) overnight at 4°C; afterward, the samples were incubated with goat anti-rabbit IgG polyclonal antibody (Abcam) or goat anti-mouse IgG polyclonal antibody (Abcam) for 1 h at room temperature. Visualization was conducted using an enhanced chemiluminescence (ECL) detection system (Thermo Fisher Scientific) using GAPDH levels as an endogenous control.

Total proteins were collected from target cells with a Cell Mitochondria Isolation Kit (Beyotime, Shanghai, China). The samples were incubated at 4 °C for one night with the primary antibodies METTL3 (15073-1-AP, Proteintech), Bcl-2 (ab196495, Abcam), Bax (50599-2-Ig, Proteintech), Caspase 9 (10380-1-AP, Proteintech), Caspase 3 (19677-1-AP, Proteintech), cleaved PARP-1 (ab32064, Abcam), E-cadherin (BF0219, Affinity), N-cadherin (AF4039, Affinity), vimentin (10366-1-AP, Proteintech), PTEN (22034-1-AP, Proteintech), YTHDF2 (ab220163, Abcam), p53 (CSB-PA07889A0Rb, CUSABIO Life Sciences, College Park, USA), c-Myc (10057-1-AP, Proteintech), NF-κb (ab76302, Abcam), Zeb1 (ab180905, Abcam), Zeb2 (ab191364, Abcam), Snail (ab180714, Abcam), Twist1 (ab50887, Abcam); after that, the samples were incubated at room temperature for 1 h with goat anti-rabbit IgG polyclonal antibody (Abcam) or goat anti-mouse IgG polyclonal antibody (Abcam). GAPDH levels were used as an endogenous control for visualization employing the enhanced chemiluminescence (ECL) detection method (Thermo Fisher Scientific).