Anti β actin mab c4

Tumor cell lines (1 × 10⁶) were washed in phosphate-buffered saline (PBS) and lysed in LDS sample buffer (Invitrogen, Thermo Fisher Scientific). The cell lysate was subjected to electrophoresis in a 4–12% NuPAGE Bis–Tris SDS-PAGE gel (Invitrogen, Thermo Fisher Scientific) under reducing condition and transferred to an Immobilon-P membrane (Merck Millipore). The membrane was blocked in PBS containing 0.01% Tween 20 and 5% non-fat dry milk for 1 h at room temperature and incubated with polyclonal rabbit anti-human PD-L1 (E1L3N, Cell Signaling Technology) diluted 1:1000 in blocking buffer for overnight at 4 °C, or anti-β-actin mAb (C4, Santa Cruz Biotechnology) diluted 1:2000 in blocking buffer as the control for 1 h at room temperature. After washing, the membrane was incubated with horseradish peroxidase-labeled sheep anti-rabbit or anti-mouse IgG and visualized using the Amersham ECL Prime Western Blotting Detection System (GE Healthcare Life Sciences).

Anti-c-Abl mAb (8E9) (catalog number: sc-56887; lot number: C2117), anti-β-actin mAb (C4) (catalog number: sc-47778; lot number: A2017), and anti-Myc/c-Myc mAb (9E10) (catalog number: sc-40; lot number: A2814) were purchased from Santa Cruz Biotechnology. Anti-HCV Core 1b mAb (clone C7-50) (catalog number: ab2740; lot number: GR3249588-2) was obtained from Abcam. Anti-mouse IgG (H+L), F(ab')2 fragment (Alexa Fluor 488 conjugate) (catalog number: 4408; lot number: 18), and anti-phospho-c-Abl (Tyr412) (247C7) (catalog number: 2865; lot number: 3) mAb were purchased from Cell Signaling Technology. Anti-HCV NS3 protein polyclonal antibodies (catalog number: GTX131276; lot number: 41836) were obtained from GeneTex. Anti-pTyr mAb (4G10) (catalog number: 05-321; lot number: 2138006) was obtained from Sigma–Aldrich.

Antibodies and their sources were as follows: anti-caveolin-1 IgG (pAb N-20), anti-c-Myc (mAb 9E10) and anti-β-actin (mAb C4) were from Santa Cruz Biotechnology (Santa Cruz, CA); anti-complex I (mAb MS112), anti-complex II (mAb MS204) and anti-complex V (mAb MS502) were from MitoSciences (Eugene, OR); anti-complex IV (mAb A21348) was from Thermo Fisher Scientific (Waltham, MA); anti-complex III (mAb ab14745) was from abcam (Cambridge, MA). All other biochemicals used were of the highest purity available and were obtained from regular commercial sources.