Goat anti mouse igg hpr

Harvested cells were lysed in RIPA buffer (150 mM sodium chloride, 1% triton X-100, 1% sodium deoxycholate, 0.1% SDS, 50 mM Tris-HCl, pH 7.5 and 2 mM EDTA) (GenDEPOT, TX, USA) containing 1% protease inhibitor cocktail (GenDEPOT). 20 μg of each sample is separated by SDS-PAGE and transferred to a nitrocellulose membrane (GE Healthcare, Chalfont St Giles, UK). Western blotting was performed as previously described by Liu et al.³⁸ with primary antibodies to mouse anti-E-cadherin 1:1000 (Cell Signalling, Danvers, MA, USA), rabbit anti-Vimentin 1:500 (Abcam, Cambridge, UK), rabbit anti-Zeb1 1:500 (Sigma), rabbit anti-IL-6 1:1000 (Abcam), rabbit anti-STAT3 1:500 (phosphor Y705: Abcam), mouse anti-STAT3 1:1000 (Cell signalling), rabbit anti-α-lamin 1:1000 and goat anti-β-actin 1:5000 (Santa Cruz Biotechnology, Santa Cruz, USA). All primary antibodies were diluted in 5% Bovine Serum Albumin (BSA) in Tris-buffered saline (TBS) containing 0.1% Tween-20 (TBST) and incubated overnight at 4 °C. Secondary antibody included goat anti-rabbit IgG-HPR 1:5000 (Santa Cruz Biotechnology), goat anti-mouse IgG-HPR 1:2000 (Santa Cruz Biotechnology) and rabbit anti-goat IgG-HPR (GenDepot, TX, USA). Protein bands were visualised using enhanced chemiluminescence reagents (Western Lighting Plus, PerkinElmer, USA).

Primary antibodies: SARS-CoV-2 Spike S1 Antibody (HC2001, GenScript Biotech, USA), ACE2 for Western blot (WB; sc-20998, Santa Cruz, USA), ACE2 for immunofluorescence (IF) (sc-73668, Santa Cruz, USA), STAT3 (12640, Cell Signalling, USA), p-STAT3 (Try705) (9145, Cell Signalling, USA), EGFR (sc-120, Santa Cruz, USA), p-EGFR(Tyr1068) (44788G, Thermo-Fisher Scientific, USA), p-EGFR(Tyr1086) (369700, Thermo-Fisher Scientific, USA), ERα (ab14020, Abcam, USA), β-actin (sc-8432, Santa Cruz, USA). Secondary antibodies: goat anti-human IgG Alexa Fluor 555 (A-21433, ThermoFisher, USA), goat anti mouse IgG-FITC (sc-2010, Santa Cruz, USA), goat anti-chicken IgY H&L Alexa Fluro 647 (ab150175, Abcam, USA), goat anti-mouse IgG H&L Alexa Fluro 647 (ab150115, Abcam, USA), goat anti-mouse IgG-HPR (sc-2031, Santa Cruz, USA), goat anti-rabbit IgG H&L-HRP (ab6721, Abcam, USA). Phalloidin labelled with Alexa Fluor 647 for F-actin staining (A22287, ThermoFisher, USA). Hoescht 33342 for nuclear (DNA) staining (H3570, ThermoFisher, USA).