N ethylmaleimide e3876

Nocodazole (M1404), PKH67 (green) (PKH67GL-1KT), PKH26 (red) (PKH26GL-1KT), brefeldin-A (B7651, used at 5 μg/ml), N-ethylmaleimide (E3876, used at 50 μM), LiCl2 (L-4408), PMA (used at 100 nM; P8139), and dynasore (D7693, used at 100 μM) were purchased from Sigma (St. Louis, MO, USA). TNF-α (210-TA) (used at 17 ng/ml), VEGF (used at 19 ng/ml; 293-VE), and neutralizing antibodies to VEGF (used at 1 μg/ml; MAB293) and VEGFR2 (used at 1 μg/ml; MAB3572) were purchased from R&D Systems (Minneapolis, MN, USA). Smac mimetic TL-32711 (A-1901) (used at 100 nM) was purchased from Active Biochem (Maplewood, NJ, USA). Cisplatin (NDC 0015-3220-22) was purchased from Bristol Laboratories (Princeton, NJ, USA). Antibodies to VEGF (for WB: Sc-7269), PKC-ζ (Sc-17781), K-Ras (Sc-30), and VEGFR2 (for WB: Sc-504) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies to E-cadherin (3195), p70S6K (2708), phospho-S6 ribosomal protein (Ser-235/236) (2211), S6 ribosomal protein (2317), syntaxin-6 (2869), and cdc42 (2466) were purchased from Cell Signaling Technology (Beverly, MA, USA). Antibody to β-catenin (610153) was purchased from BD Transduction Laboratories (Lexington, KY, USA). BI-D1870 (used at 5 μM; S2843) was purchased from Selleckchem (Houston, TX, USA).

SD70 (M60194-2s) was purchased from Xcessbio Biosciences Inc; Cycloheximide (CHX; N11534), Calpeptin (C8999), chloroquine (C6628), iodoacetamide (I1149), and N-ethylmaleimide (E-3876) were purchased from Sigma-Aldrich; Sorafenib (8705) was purchased from Cell Signaling Technology; MG-132 (S2619) was purchased from Selleck; Z-VAD-FMK (2163) was purchased from Tocris. Treatment conditions were described in detail in the text.

Antibodies against Flag (F3165, 1:2000) and HA (H6908, 1:5000) were purchased from Sigma (Burlington, MA, USA). Antibodies against the envelope protein of ZIKV (GTX133314, 1:1000), NS1 protein of ZIKV (GTX133307, 1:1000), glyceraldehyde-3-phosphate dehydrogenase (GAPDH; GTX100118, 1:5000), and ZDHHC11 (GTX106800, 1:1000) were purchased from GeneTex (Hsinchu City, Taiwan, P.R.C). For the immunofluorescence studies, antibodies against the envelope protein of flaviviruses (ab214333, 1:100) were purchased from Abcam (Shanghai, China). N-ethylmaleimide (E3876), hydroxylamine hydrochloride (255580), and 2-bromopalmitate (2-BP; 21604) were obtained from Sigma (Burlington, MA, USA). BMCC-biotin (21900) was purchased from Thermo Fisher Scientific (Waltham, MA, USA). Streptavidin-horseradish peroxidase (M00091) was purchased from Genscript (Piscataway, NJ, USA). Fluorescein (FITC)-conjugated AffiniPure goat anti-mouse immunoglobulin (Ig)G (H+L) (SA00003-1) and Cy3-conjugated AffiniPure goat anti-rabbit IgG (H+L) (SA00009-2) were purchased from ProteinTech (Rosemont, IL, USA).