Antibody against vinculin

Cultured cells were fixed with 1% paraformaldehyde in PBS for 30 min and permeabilized by treatment with 0.05% Triton X-100 in PBS for 5 min. The fixed cells were incubated with 10% normal goat serum for 30 min at room temperature and then stained with an antibody against vinculin (Sigma, St. Louis, MO, USA) as a marker of focal adhesions for 60 min. After washing in PBS for 20 min, the fixed specimens were incubated with fluorescein-conjugated anti-mouse IgG. Samples were then observed by conventional epifluorescence microscopy or phase-contrast microscopy (Olympus, Tokyo, Japan).

Cisplatin (S1166), quisinostat (S1096), paclitaxel (S1150), and topotecan (S9321) were purchased from Selleck Chemicals. Cyclophosphamide (CA80500-080), histamine (H7125), and thapsigargin (T9033) were from Sigma-Aldrich. Mitotracker deep red FM was from Thermo Fisher Scientific (M22426). Antibodies against calregulin (Cat# sc-166837), HSP90 (Cat# sc-13119), and β-Actin (Cat# sc-47778) were from Santa Cruz Biotechnology; antibodies against SMARCA2 (Cat# 11996), cleaved PARP (Cat# 5625), and cleaved caspase 3 (Cat# 9664) were from Cell Signaling; antibodies against MICU2 (Cat# ab-101465), VDAC1 (Cat# ab-14734), and GRP75 (Cat# ab-2799) were from Abcam; antibody against SMARCA4 (Cat# A300-813A) was from Bethyl Laboratories (A300-813A); antibody against IP3R3 (Cat# 610312) was from BD Pharmingen; antibody against vinculin (Cat# V4505) was from Sigma-Aldrich; antibody against MCU (Cat# HPA0168480) was from Atlas; and antibody against MICU1 (Cat# orb-323178) was from Biorbyt. Antibody against SMARCA4 was used with 1:5000 dilution and all others with 1:1000 dilution. Antibodies for IHC are described in the Immunohistochemistry method section below. Secondary antibodies goat anti-mouse IgG (Cat#1706516) and goat anti-rabbit IgG (Cat#1706515) were purchased from Bio-Rad.

5-FU was purchased from Calbiochem (San Diego, CA, USA). Curcumin, X-treme GENE™ HP DNA transfection reagent and antibody against vinculin were procured from Sigma-Aldrich (St. Louis, MO, USA). Antibodies against p-ERK1/2, p-p38, p-JNK, p-Akt, p-IKK, c-Myc and caspases were obtained from Cell Signaling Technology (Beverly, MA, USA). Antibodies against p-p65, MDR-1, VEGF, Bcl-2, cIAP1, XIAP, survivin, TS, PARP, Cox-2 and GAPDH were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibody against ABCG2 was purchased from Abcam (Cambridge, Mass, USA).