Ab233389

For immunoblotting, proteins were isolated from cells using RIPA buffer. Total protein extracts (15–50 μg) were separated using sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred onto polyvinylidene fluoride membrane. Membranes were then probed with anti-bax (1:2000; #ab32503; Abcam), bcl-2 (1:2000; #ab196495; Abcam), cleaved-caspase-3/caspase-3 (1:2000; #ab184787; Abcam), SM22α (1:2000; #ab14106; Abcam), calponin (1:500; #ab227661; Abcam), SMMHC (1:2000; #ab125884; Abcam), vimentin (1:2000; #ab92547; Abcam), collagen I (1:1000; #ab270993; Abcam), osteopontin (OPN; 1:1000; #ab63856; Abcam), LAMC1 (1:1000; #ab233389; Abcam), EVI1 (1:1000; #SAB2100723; Sigma), AKT3 (1:2000; #ab152157; Abcam), TP53INP1 (1:2000; #ab202026; Abcam), and β-actin (1:2000; #ab8226; Abcam) at room temperature for 1.5 h. Then, membranes were immersed with the HRP-conjugated secondary antibody at room temperature for 1 h. Following this, the BM chemiluminescence blotting system (Thermo Scientific) was used for detection and protein bands were quantified using Image J software (NIH, USA).

After protein quantization, 15 μg of protein was loaded, resolved on 10% sodium dodecyl sulfate (SDS)-polyacrylamide gels, electrotransferred onto nitrocellulose membranes, and blocked with serum. The membranes were incubated overnight with rabbit antibodies against ITGA1 (1:1000 dilution; ab181434; Abcam, Cambridge, UK), rabbit antibodies against ITGB1 (1:5000 dilution; ab183666; Abcam, Cambridge, UK), rabbit antibodies against LAMC1 (1:1000 dilution; ab233389; Abcam, Cambridge, UK), and rabbit antibodies against CKM (1:1000 dilution; sc-365046; Santa Cruz Biotechnology, Inc., Dallas, TX, USA). After 3 times of washing, secondary antibodies were added. Blots were detected by an enhanced chemiluminescence (ECL) reagent (ECL-Plus/Kit; Amersham, Piscataway, NJ, USA). Besides, β-actin (1:1000 dilution; Sigma-Aldrich, St. Louis, MO, USA) was used as loading control.