Sc 33684

The following antibodies were used for western blotting: rabbit monoclonal anti-CD24 (1:500, catalog number ab179821, Abcam, MA, USA), rabbit monoclonal anti-CD44 (1:500, catalog number ab189524, Abcam), rabbit recombinant multiclonal anti-CD133 (1:800, catalog number ab278053, Abcam), mouse monoclonal anti-Nanog (1:1000, catalog number ab173368, Abcam), rabbit monoclonal anti-OCT4 (1:500, catalog number ab200834, Abcam), mouse monoclonal anti-SOX2 (1:1000, catalog number ab79351, Abcam), rabbit polyclonal anti-DLG5 (1:1000, catalog number ab231283, Abcam), rabbit polyclonal anti-PRDM16 (1:1000, catalog number ab106410, Abcam), mouse monoclonal anti-ErbB-2 (1:500, catalog number sc-33684, Santa Cruz Biotechnology, CA, USA), and mouse monoclonal anti-β-actin (1:1000, catalog number A5441, Sigma-Aldrich, MO, USA). Other primary antibodies and secondary antibodies and experimental procedures for immunoblotting are provided in the supplementary experimental procedures^{10 (link)}.

The expression of EGFR, HER2, and E-cadherin on the surface of the oral epithelial cells was quantified by flow cytometry. Briefly, OKF6/TERT-2 cells in 6-well tissue culture plates were incubated with tissue culture medium with or without IFN-γ for 24 h and then infected with 5 × 10⁵C. albicans cells. After 75 min, the cells were scraped from the wells with a cell scraper, fixed with 3% paraformaldehyde, blocked with 1% goat serum, and then stained with specific antibodies (for EGFR, sc-101, and for HER2, sc-33684, from Santa Cruz Biotechnology; for E-cadherin, ab1416 from Abcam, Inc.), followed by an Alexa 488-conjugated goat or mouse anti-rabbit antibody (Life Technologies, Inc.). Control epithelial cells were incubated in a similar concentration or mouse or rabbit IgG (Abcam, Inc.). The fluorescence of the cells was determined by flow cytometry, analyzing at least 10,000 cells per condition.

For histology, tissues were fixed in the 10% formalin, blocked in paraffin, sectioned as 5 µm thick, stained with hematoxylin and eosin, and examined by light microscopy. Antibodies against ERalpha (sc-542; Santa Cruz), PR (sc-166169; Santa Cruz), ERBB2 (sc-33684; Santa Cruz), and MRC2 (sc-271148; Santa Cruz) were used for immunohistochemical staining, by using a Histostain® Plus Broad Spectrum kit (859043; Life Technologies) as per the manufacturer's instruction. Antibodies against Keratin14 (PRB-155P; Covance), Keratin18 (sc-53256; Santa Cruz), E-Cadherin (3195; Cell Signaling Technology) and Vimentin (5741; Cell Signaling Technology) were used for immunoflouresencent staining, and nuclei were stained with DAPI (62248; ThermoFisher Scientific).