Goat anti β actin sc 1615

Cells were inoculated at a density of 7 × 10⁴/well on 6-well plates and cultured for 96 h. The cells were collected in cold phosphate-buffered saline and precipitated by centrifugation. Cell lysates were prepared as described previously [17 (link)]. The aliquots (30–50 μg) were applied to SDS-polyacrylamide gel electrophoresis and Western blotting. Rabbit anti-AMD1 (11052-1-AP), anti-GSK3β (22104-1-AP), and anti-SRM (15979-1-AP) antibodies were obtained from ProteinTech (Tokyo, Japan). Goat anti-β-actin (sc-1615) and rabbit anti-p-ERK1/2 (sc-16982-R) antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Rabbit anti-ERK1/2 (#4695) and anti-p-GSK3β (#5558) antibodies were from Cell Signaling Technologies (Beverly, MA, USA). These primary antibodies were used at the dilution 1:1000. The blotted membrane was scanned with a C-DiGit Blot Scanner (LI-COR Biotechnology, Lincoln, NE, USA).

Mouse anti-ZO-1 monoclonal antibody (33–9100, Lot: 1100420A) and Lipofectamine 2000 were obtained from Thermo Fisher Scientific (Rockford, IL, USA). Rabbit anti-p-Akt (4060, Lot: 14), anti-Akt (4691, Lot: 20), anti-p44/42 MAPK (ERK1/2, 4695, Lot: 1), anti-c-Fos (2250, Lot: 4), and anti-MRP2 (ABCC2, 4446, Lot: 1) polyclonal antibodies were from Cell Signaling Technology (Beverly, MA, USA). Goat anti-β-actin (sc-1615, Lot: C141), rabbit anti-p-ERK1/2 (sc-16982R, Lot: L0704) polyclonal antibodies and mouse anti-p-c-Fos (sc-81485, E2008) monoclonal antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Rabbit anti-ABCB1 (GTX108354, Lot: 39834), anti-ABCC1 (GTX116046, Lot: 40135), and anti-ABCG2 (GTX100437, Lot: 39471) polyclonal antibodies were from GeneTex (Irvine, CA, USA). Mouse anti-FLAG (018-22381, Lot: SAQ1191) monoclonal antibody, CDDP, and DXR were from Wako Pure Chemical (Osaka, Japan). LY-294002, and U0126 were purchased from BIOMOL Research Laboratories (Plymouth Meeting, PA, USA) and Sigma-Aldrich (Saint Louis, MO, USA), respectively. All other reagents were of the highest purity commercially available.