Anti ppm1a

SCC25 and FADU cells were harvested and lysed in RIPA Lysis Buffer (Yita Biology, Pinggu, Beijing, China) to collect proteins. Proteins were separated using SDS-PAGE at 30 μg/lane and transferred to a nitrocellulose membrane (EMD Millipore, Pudong, Shanghai, China). Next, our group blocked the membrane with 5% dried skimmed milk for 1 h. Then, the membrane was incubated with primary antibodies including anti-PPM1A (Cat no. ab14824, Abcam) and anti-GAPHD (Cat no. ab8245; Abcam) at 4°C throughout the night and with horseradish peroxidase- (HRP-) conjugated secondary antibodies (Guduo, Shanghai, China) for 1 h under ambient temperatures. The researchers employed ImageJ software for quantifying the density exhibited by the respective band.

Specific antibodies were purchased from the following commercial sources: Anti-AFP, anti-ALB, anti-CD44, anti-Evi1, anti-flag (mouse), anti-HA, anti-HNF4α, anti-H3, anti-H3K36me3, anti-Myc, anti-OCT4A, anti-P300, anti-PPM1A (rabbit), anti-pSmad2 (S465/467), anti-pSmad2 (S245/250/255), anti-pSmad3 (s423/425), anti-Smad2, anti-Smad3 (rabbit), anti-SnoN, anti-Sox2, anti-TAT, and normal rabbit IgG from Cell Signaling Technology (Danvers, MA); anti-PPM1A (mouse), anti-SC35, and anti-SETD2 were from Abcam (Cambridge, MA); Anti-Smad4 and normal mouse IgG were from Santa Cruz Biotechnology (Santa Cruz, CA); Anti-β-actin, and anti-flag (rabbit) from Sigma-Aldrich (St. Louis, MO); Alexa594 goat anti-mouse IgG from Life Technology (Carlsbad, CA); Dylight488 goat anti-rabbit IgG from Vector Labs (Burlingame, CA).