Sc 73668

Western blot analyses were performed according to Melamed et al. 2004 [21 (link)] with the following modifications: nitrocellulose membranes (iBlot 2 Transfer Stacks; IB23001, Invitrogen, Carlsbad, CA, USA) were incubated with mouse monoclonal anti-ACE2 (AC18Z) (sc-73668, Santa Cruz, 1:200 dilution) overnight at 4 °C. Detection of the primary antibodies was with a secondary, horseradish peroxidase (HRP)-conjugated goat anti-mouse antibody (115-035-003, Jackson ImmunoResearch, 1:10000 dilution) for 1 h at room temperature, and with Immobilon Forte Western HRP substrate (WBLUF0500, Millipore).

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).