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Rabbit polyclonal anti-cortactin antibodies were obtained from Santa Cruz Biotechnology (Dallas, TX, USA). Alexa Fluor 568-conjugated phalloidin, secondary anti-rabbit antibodies conjugated with Alexa Fluor 488, gelatin conjugated with FITC, fetal bovine serum (FBS), trypsin, glutamine, penicillin/streptomycin, and DMEM were purchased from Invitrogen (Carlsbad, CA, USA). Dako fluorescent mounting medium was obtained from Dako (Agilent, Santa Clara, CA, USA). The epidermal growth factor (EGF) and rat-tail collagen (acid-extracted, non-pepsin treated) type I were obtained from BD Biosciences (Agilent, Santa Clara, CA, USA). Cell Proliferation Kit II was purchased from Roche (Basel, Switzerland). Antibodies directed against EGFR (SC-03), MET (SC-10), AKT1/2/3 (SC-8312), and phospho-AKT1/2/3 (S473; SC-135651) were purchased from Santa Cruz Biotechnologies. Anti-phospho-EGFR (Y1069; catalogue no. 3777), anti-phospho-MET (Y1234/Y1235; no. 3077), anti-ERK1/2 (no. 9102), and anti-phospho-ERK1/2 (T202/Y204; no. 9101) antibodies were from Cell Signaling Technologies (Danvers, MA, USA). HGF was obtained from Sigma Aldrich (Saint Louis, MO, USA), foretinib from Santa Cruz Biotechnologies, and lapatinib from Selleckchem (Houston, TX, USA). All other chemicals were classified as analytical grade reagents.

The antibodies used were as follows: anti-Banf1 N-terminus (SAB1409950, Sigma-Aldrich and ab88464, Abcam, 1:1000 for WB and 1:500 for IF), anti-Banf1 C-terminus (PRS40170604, Sigma-Aldrich, 1:1000 for WB and 1:500 for IF), PARP1 (9532, Cell Signalling Technology 1:1000 for WB and 1:500 for IF), PARP1 (ab191217, Abcam Fig. 2e 1:500 for IF), anti-Emerin (5430, Cell Signalling Technology, 1:500 for IF), anti-Flag M2 Antibody (F3165, Sigma-Aldrich, 1:1000 for WB and 1:300 for IF), anti-PAR (ab14459, Abcam, 1:1000 for WB), anti-γ-Tubulin (T6557, Sigma-Aldrich, 1:2000 for WB), anti-H3 (4499, Cell Signalling Technology, 1:2000 for WB), anti-H4 (2935 Cell Signalling Technology 1:1000 for WB), anti-SP1 (9389, Cell Signalling Technology, 1:1000 for WB), anti-EGFR (sc-03, Santa-cruz, 1:500 for WB), anti-LC3B (2775, Cell Signalling Technologies, 1:1000 for WB), anti-phospho-p53 ser15 (2984, Cell Signalling Technology, 1:1000 for WB), anti-β-actin (612656, BD Biosciences, 1:2000 for WB). Fluorescent secondary antibodies used were: Donkey anti-Mouse 800 nm (LiCor; IRDye 800CW 926-32212, 1:5000 for WB), Donkey anti-Rabbit (LiCor; IRDye 680LT 926-28023, 1:5000 for WB) and Alexa Fluor 488 (Cat# A32766, Molecular Probes, 1:200 for IF) and 594 (Cat# A32754, Molecular Probes, 1:200 for IF).

H292, H292-HPV16E5, H292-HPV16E6, and H292-HPV16E7 cells were stimulated with 100 ng/mL epidermal growth factor (EGF, Sigma-Aldrich, St. Louis, MO, USA). They were incubated at 37 °C for 30 min. Then, nuclear and non-nuclear fractions were isolated and subjected to Western blotting with anti-EGFR (sc-03, Santa Cruz, Dallas, TX, USA) and anti-pEGFR (sc-12351R, Santa Cruz) antibodies on nitrocellulose membranes. The same membranes were also probed with histone 3 (Histone H3 Antibody #9715, Cell Signaling Technology, for the nuclear fraction) or tubulin (α-Tubulin Antibody #2144, Cell Signaling Technology (Danvers, MA, USA), for the non-nuclear fraction) as a loading control.

10⁵ cells were seeded onto cover slides in six-well plate. The next day, the cells were rinsed with Dulbecco’s Phosphate-Buffered Saline (DPBS, Thermo Fisher Scientific) and then fixed with 4% paraformaldehyde (Sigma-Aldrich) for 10 min. at room temperature, followed by permeabilization with 0.1% Triton X-100 (Sigma-Aldrich). The cells were stained with anti-pan CK (1:100, NB120-6401, Novus biological), anti-EGFR (1:30, sc-03, Santa Cruz) or anti-α-smooth muscle actin (α-SMA, 1:200, ab5694, Abcam) antibody overnight at 4 °C. The cells were then washed with DPBS three times for 10 min. per wash and then incubated with DyLight 488-labeled (1:500, 611-141-002, Thermo Fisher Scientific) or DyLight 549-labeled secondary antibody (1:500, 611-142-002, Thermo Fisher Scientific) at room temperature for one hour. After washed with DPBS three times for 10 min each, cover slips were counterstained with 4′,6-diamidino-2-phenylindole (DAPI, Vector Lab, Burlingame, CA, USA), mounted and viewed under fluorescent microscopy.