Anti lamin b1 antibody

Total cell lysates and cytoplasmic and nuclear protein fractions were extracted from cell cultures as described previously [51 (link)]. Standard Western blot analysis of protein expression was carried out using primary anti-HA (rabbit; Cell Signaling Technology), anti-TAZ (rabbit; Cell Signaling Technology), anti-YAP (rabbit; Cell Signaling Technology), anti-E-cadherin (mouse; BD Biosciences), anti-vimentin (rabbit; Cell Signaling Technology), anti-Merlin (rabbit; Santa Cruz Biotechnology), anti-LATS1 (rabbit; Cell Signaling Technology), anti-MST1 (rabbit; Cell Signaling Technology), anti-phosphorylated LATS1 (Thr1079, rabbit; Cell Signaling Technology), anti-phosphorylated MST1 (Thr183)/MST2 (Thr180, rabbit; Cell Signaling Technology), anti-TEAD (rabbit; Cell Signaling Technology), and anti-CTGF (mouse; Santa Cruz Biotechnology) antibodies. Equal protein-sample loading was monitored using an anti- glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody for total cell protein lysates (rabbit; Santa Cruz Biotechnology), an anti-α-tubulin antibody for cytoplasmic fractions (mouse; Oncogene), and an anti-lamin B1 antibody for nuclear fractions (goat; Santa Cruz Biotechnology).

Protein extracts (40 μg each) were separated using 12% (wt./vol.) SDS‐PAGE, transferred onto a polyvinylidene fluoride (PVDF) membrane and probed with respective antibodies for ACAT2 (Abcam), COL1A1 and COL3A1 (Proteintech, Chicago, IL, USA), p‐Smad3, Smad3 (Cell Signaling Technology, Beverly, MA, USA) overnight at 4°C. The membranes were then washed extensively with TBS/T and incubated with a horseradish peroxidase (HRP)‐conjugated secondary antibody (Santa Cruz) for 1 hour at room temperature. The protein was visualized using the ECL Plus detection system (GE Healthcare, WI, USA). As internal controls, the membranes were also immunoblotted with anti‐GAPDH and anti‐Lamin B1 antibody respectively (Santa Cruz).

MeHg and avidin-agarose was obtained from Sigma-Aldrich (St. Louis, MO, USA). Escherichia coli BL21 cells and trypsin were purchased from Promega Co. (Madison, WI, USA). Anti-GAPDH antibody and anti-Lamin B1 antibody were obtained from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA). Anti-G6PD antibody, BPM, 4,6-diamidino-2-phenylindole (DAPI), annexin V fluorescein isothiocyanate (FITC), and propidium iodide were obtained from Bethyl Laboratories (Montgomery, TX, USA), Dojindo (Kumamoto, Japan), Nacalai Tesque Inc. (Kyoto, Japan), MBL (Aichi, Japan), Wako Pure Chemical Industries (Osaka, Japan), respectively. Anti-Akt, anti-CREB, anti-phosphorylated Akt (Ser473), anti-phosphorylated CREB (Ser133), anti-Bcl-2, horseradish peroxidase (HRP)-conjugated anti-rabbit antibodies, LY294002, and wortmannin were purchased from Cell Signaling Technology (Beverly, MA, USA). Alexa 488-secondary antibody, rhodamine phalloidin and Lipofectamine 2000 were obtained from Thermo Fisher Scientific (MA, USA). All other reagents used were of the highest purity available.