Mouse anti human gapdh antibody

Total protein was isolated from cells using cell lysis buffer (Cell Signaling Technology, Inc.) after transfection for 48 h. The protein levels were quantified using a DC Protein Assay (Bio-Rad Laboratories, Hercules, CA, USA). Protein samples (30 μg) were loaded on a 12% SDS-PAGE gels and electroblotted to Immun-Blot polyvinylidene fluoride membranes (Millipore, Billerica, MA, USA). Membranes were blocked and probed with a monoclonal rabbit anti-human Bcl-xL antibody (1:1,000; Epitomics Inc, Burlingame, CA, USA), then washed with Tris-buffered saline and Tween 20 (50 mM Tris, 150 mM NaCl, 0.1% Tween-20; pH 7.6; Sigma-Aldrich), and incubated with a secondary horseradish peroxidase-conjugated goat anti-rabbit antibody (1:5,000; Hangzhou Hua’an Biotechnology Co., Ltd., Hangzhou, China). Protein levels were normalized to total glyceraldehyde 3-phosphate dehydrogenase (GAPDH) using a mouse anti-human GAPDH antibody (1:1,000; Abcam, Cambridge, UK). The intensity of each protein band was quantified by Quantity One version 4.62 software (Bio-Rad Laboratories).

Rabbit anti-human LAMA1 antibody (sc-5582) was from Santa Cruz Biotechnology (Dallas, TX, USA). Mouse anti-human GAPDH antibody, goat anti-rabbit IgG H&L (ab150077), and goat anti-mouse IgG H&L were purchased from Abcam (Cambridge, MA, USA). TRIzol reagent and DAPI (4′,6-diamidino-2-phenylindole) were obtained from Invitrogen (Carlsbad, CA, USA). The PrimeScript RT reagent kit with gDNA Eraser and SYBR Premix Ex Taq™ II (Tli RNaseH Plus) kit were from Takara Clontech (Kyoto, Japan). Cell Counting Kit-8 (CCK-8) was from Dojindo (Kumamoto, Japan), and 4-well culture inserts were from Ibidi (cat. number 80469; Martinsried, Germany).