Anti actin 8h10d10

RIPA lysis buffer was used to extract protein from cell lysis. Anti‐Tubulin mouse (T5168) was acquired from Sigma (Wuxi City, China). Anti‐actin (8H10D10) was acquired from Cell Signaling Technology (Beijing, China). Anti‐RNF168 (SC‐101125) was from Santa Cruz Biotechnology (Shanghai, China). Anti‐STAT1 (9172) was acquired from Cell Signaling Technology. anti‐Lamin B1 (66095‐1) was acquired from Wuhan Sanyin company (Wuhan, China).

Total cell extracts were prepared by resuspending 3 × 10⁶ NK cells in 100 μL NP-40 lysis buffer (50 mM Tris-HCl, pH 7.5, 120 mM NaCl, 20 mM NaF, 1 mM EDTA, 6 mM EGTA, 15 mM sodium pyrophosphate, 1 mM PMSF, 0.1% Nonident P-40). Fifteen minutes of cell lysis on ice was followed by centrifugation for 20 min at 14,000 × g. Cleared lysates were analyzed directly by SDS-PAGE and Western Blotting. Briefly, equal amounts of protein were separated by SDS-PAGE, transferred to nitrocellulose membranes (Thermo Fisher), blocked in 5% dry milk powder dissolved in 1×PBS-T, and then probed with primary antibody and HRP-conjugated secondary antibody (Santa Cruz Biotechnology, Dallas, TX, USA). Proteins were visualized using Enhanced Chemiluminescent solution (Thermo Fisher) and FUSION Vilber imager (Eberhardzell, Germany). The intensity of signals was quantified by densitometric analysis using the image analysis software ImageJ (Version 1.51j8). The value for HIF-1α was normalized to that for β-Actin. Anti-HIF-1α (# 2185) was obtained from Abcam (Cambridge, UK) and Anti-Actin (8H10D10) from Cell Signaling Technology (Frankfurt am Main, Germany). Representative experiments out of three performed are shown.

Cells were lysed with RIPA lysis buffer. Anti-ERα (HC-20, SC543) was from Santa Cruz Biotechnology. Anti-SHARPIN (AB69507), and anti-FLAG (M2, ab48763) were acquired from Abcam. Anti-actin (8H10D10) was acquired from Cell Signaling Technology.

Cells were lysed with RIPA lysis buffer. Anti- ER alpha mouse (1D5, SC56833) was from Santa Cruz Biotechnology. Anti-ER alpha rabbit (D8H8, #8644) was from Cell Signaling Technology. Anti-SMURF1 (AB117552), anti-Myc mouse (AB32), anti-Myc rabbit (AB9106) and anti-FLAG (M2, ab48763) were acquired from Abcam. Anti-HA (MMS-101R) was acquired from COVANCE. Anti-actin (8H10D10) was acquired from Cell Signaling Technology.