Anti caveolin 1

Ammonium chloride (NH₄Cl) and chloroquine were purchased from Sigma (St. Louis, MO, USA). Chlorpromazine (CPZ), methyl-β-cyclodextrin (MβCD), dynasore, and wortmannin were purchased from TargetMol (Boston, MA, USA). 5-Ethyl-N-isopropyl amiloride (EIPA) was purchased from MedChemExpress (Monmouth Junction, NJ, USA).
An anti-FAdV-4-fiber2 monoclonal antibody (mAb 3C2) was kindly provided by Prof. Jianqiang Ye (Yangzhou University, Yangzhou, Jiangsu, China). A rabbit polyclonal anti-FAdV-4-penton antibody was generated by our laboratory. Rabbit polyclonal anti-caveolin-1 and clathrin HC antibodies were purchased from Santa Cruz Biotechnology (CA, USA). Anti-β-actin antibodies, HRP-conjugated secondary antibodies, and FITC-conjugated anti-mouse and anti-rabbit IgG were purchased from TransGen Biotech (Beijing, China). TRITC-phalloidin and FITC-dextran were purchased from Sigma.
A pGFP-V-RS (vector), scramble shRNAs, and shRNAs targeting clathrin heavy chain (CHC) (shCHC) were kindly provided by Dr Hung-Jen Liu (Institute of Molecular Biology, National Chung Hsing University, Taichung, Taiwan). siRNA targeting caveolin-1 (5′-CCACTTTCACTGTAACAAA-3′) and negative siRNA were synthesized by RiboBio (Guangzhou, China).

Anti- Signal transducer and activator of transcription-3 (STAT3), anti-TSHR, anti-Caveolin 1, fluorescein isothiocyanate (FITC)-conjugated secondary antibody were obtained from Santa Cruz Biotechnology, Inc. (California, USA); SDS-PAGE molecular weight standards were purchased from Bio-Rad Laboratories (Hercules, CA, USA). Chemiluminescence kits was purchased from Amersham (Rainham, Essex, UK). Cholesterol (CHO), TSH and cAMP EIA kits were purchased from Sigma-Aldrich Corporation (St. Louis, MO, USA) and CABRU SAS (Milan, Italy), respectively. Sphingomyelin (SM) 18∶1 12∶0, SM 18∶1 16∶0, SM 18∶1 18∶1, SM 24∶0, phosphatidylcholine (PC) 16∶0 18∶1, PC 16∶0 20∶4, PC 18∶1 18∶0, ceramide 18∶1 16∶0, ceramide 18∶1 18∶0, ceramide 18∶1 20∶0, ceramide 18∶1 24∶0 were purchased from Avanti (Avanti Polar, Alabaster, USA).

Cells (1 × 10⁶) were seeded in 25 cm² flasks and exposed or not to radiation. Thirty minutes after irradiation, cells were harvested, rinsed, and lysed in RIPA buffer (Santa Cruz Biotechnology, Inc., Heidelberg, Germany). Cytosolic protein fractions and total lysates (30 µg of each) were separated using SDS-PAGE (12% polyacrylamide gels) and electrotransferred onto nitrocellulose membranes (Bio-Rad, Marnes-la-Coquette, France). Membranes were then incubated with anti-p-AKT (S473), AKT-pan (Cell Signaling Technology, Danvers, MA, USA), anti-flotillin-1, and anti-caveolin-1 (Santa Cruz Biotechnology, Inc., Heidelberg, Germany) primary antibodies at dilution 1:1000. Secondary antibodies were horseradish peroxidase-conjugated anti-mouse or anti-rabbit (Cell Signaling Technology), and proteins were detected using the ECL detection system Azure c300 (Azure Biosystems, Inc., Dublin, CA, USA).

N6-Isopentenyladenosine (i6A), 3-(N-Morpholino)propanesulfonic acid (MOPS) and NaCl-Sucrose were purchased from Sigma-Aldrich, Inc, St. Louis, MO, USA. The antibodies used: anti-Rap1A, anti-RAS, anti-caspase 3 (total and cleaved forms), anti- PARP (total and cleaved forms) were from Abcam, Cambridge, UK; anti-Caveolin-1 was from Santa Cruz Biotechnology, Dallas, TX, USA; anti-GAPDH and horseradish peroxidase-conjugated secondary antibodies were purchased from Cell Signaling Technologies, Danvers, MA, USA.

Dehydroisoandrosterone 3′-sulphate (cat no. D5297), hematoxylin (cat no. H3136), anti-beta-actin (cat no. A2668) and goat anti-Mouse IgG (γ-chain specific)-HRP (cat no. A3673) were purchased from Sigma Aldrich Inc., St Louis, MO, USA. Immobilon-P PVDF membrane (0.45 μm), ECL reagent kits (cat no. WBKLS0500), Protein-A-Agarose suspension (cat no. IP02) and goat anti-rabbit-HRP IgG (cat no.621140380011730) were procured from Merck-Millipore, Cedex, France. Other primary antibodies against phospho-Vav (Y174) (cat no. ab47282), phospho-Rac1/Cdc42 (S71) (cat no. ab5482) and Rac1 (cat no. ab33186) were purchased from Abcam, Cambridge, MA, USA. Anti-Vav (cat no. sc132) and anti-Caveolin1 (cat no. sc894) were purchased from Santa Cruz Biotechnology, CA, USA. Non-fat milk (cat no.170-6404) and precision plus protein standard marker (cat no. 161-0374) were obtained from Bio-Rad Lab., Inc., Hercules, CA., USA. Protein assay kit (cat no. 23225) was procured from Thermo-Scientific, Rockford, USA. The G-LISA Rac1 activation assay Biochem Kit (cat no. BK128) was purchased from Cytoskeleton, Denver, CO, USA. Inhibin B Enzyme Immunoassay kit (cat no. EIA-INB-1) was purchased from RayBiotech, Inc., Norcross, GA, USA. 17 β-estradiol assay kit (cat no. ADI-900-008) was obtained from Enzo Life Science, Inc., Farmingdale New York, USA.

Anti-neu/HER2 (ErbB2) antibody was from Invitrogen. Anti-ErbB2, anti-ErbB3, anti-EGFR, anti-caveolin-1, anti-phospho-ERK(Tyr204), anti-ERK, anti-phospho-Stat3(Tyr705), and anti-NDRG1 antibodies were from Santa Cruz Biotechnology INC. Anti-phospho-EGFR(Tyr1068) and anti-phospho-ErbB2(Tyr1248) antibodies were from Abcam. Anti-α-tubulin, anti-β-actin, and anti-cholera toxin B subunit antibodies were from Sigma Aldrich. Anti-4EBP1, anti-AMPKα, anti-acetyl-CoA carboxylase, anti-S6 ribosomal protein, anti-p70 S6 kinase, anti-Akt, anti-phospho-4EBP1(Thr37/46), anti-phospho-Akt(Thr308), anti-phospho-Akt(Ser473), anti-phospho-acetyl-CoA carboxylase(Ser79), anti-phospho-AMPKα(Thr172), anti-phospho-p70 S6 kinase(Thr 389), and anti-phospho-S6 Ribosomal protein(Ser240/244) antibodies were from Cell Signaling. Anti-Sestrin2 and anti-REDD1 antibodies were from Proteintech. Cholera toxin subunit B (CT-B) Conjugates were from Molecular Probes. Cy^TM3 Conjugate was from Jackson ImmunoResarch Lab. EGF was from Perprotech. MEDICA [α,α′-tetramethyl hexadecanedioic acid, HOOC-C(CH3)2-(CH2)12-C(CH3)2-COOH] was synthesized as previously described [17 (link), 26 ].