Journal: Virology journal
Article Title: SARS-CoV-2 viral protein Nsp2 stimulates translation under normal and hypoxic conditions.
doi: 10.1186/s12985-023-02021-2
Figure Lengend Snippet: Fig. 7 Nsp2 may affect composition of the cap-binding complexes. (A) Left, the representative images of western-blot analyses of proteins expressed in control and Nsp2-cells under normal and hypoxic conditions. Arrow indicates position of Strep-Nsp2. Right, ratio of densitometric signal of P-4EBP to total 4EBP in cells grown under normal or hypoxic conditions normalized to that in control cells grown under normal conditions, set as one (± SD, n = 3). (B) Left, the representative images of western blot analysis of proteins bound to m7GTP-Sepharose under normal and hypoxic conditions in control and Nsp2-cells. Arrow indicates position of Strep-Nsp2. Right, ratios of densitometric signals of eIF4G (upper graph) and eIF4E2 (lower graph) to eIF4E bound to m7GTP-Sepharose in experiments presented in lanes 1–4 (± SD, n = 3). Graph eIF4E/eIF4G: p = 0.014 for C–H versus C–N; p = 0.033 for Nsp2-H versus C–H. Graph eIF4E2/eIF4E: p = 0.03 for C–H versus C–N; p = 0.025 and 0.023 for Nsp2–H versus C–N and versus C–H, respectively
Article Snippet: Rabbit monoclonal anti-4EHP (eIF4E2) Cell signaling Technology Cat# 6916, RRID: AB_10839268 Mouse monoclonal anti-FGF2 Santa Cruz Biotechnology Cat# 145 Rabbit monoclonal anti-GAPDH Cell signaling Technology Cat# 2118, RRID: AB_561053 Mouse anti-strep Invitrogen Cat# MA5-17,283 Rabbit anti-eIF4E2 Invitrogen Cat# PA5-110849 Rabbit Calnexin Invitrogen Cat# PA5-34754 Rabbit RbS3 Cell signal tech Cat# 2579S Goat anti-Mouse Alexa FluorTM 488 Life Technologies Cat# A11029 Goat anti-Rabbit Alexa FluorTM 594 Life Technologies Cat# A11012 Rabbit polyclonal antiP-mTOR (S2448) Cell signaling Technology Cat# 2971,RRID: AB__330970 Rabbit monoclonal anti-mTOR Cell signaling Technology Cat# 2983,RRID: AB_ 2105622 Rabbit polyclonal antiP-4EBP(S65) Cell signaling Technology Cat# 9451,RRID: AB_330947 Rabbit polyclonal anti4EBP (PHAS-1) Invitrogen Cat# 51-2900 Rabbit monoclonal anti-P-eIF2α (Ser51) Cell signaling Technology Cat#9721,RRID: AB_330951 rabbit monoclonal anti-eIF2α(D7D3) XP Cell signaling Technology Cat#9721,RRID: AB_10692650 Mouse monoclonal anti-eIF4E2 (Figs. 6, 7, and Additional file 1: Figure S1) Invitrogen Cat#MA5-25412 Mouse monoclonal anti-GAPDH (Fig. 5C and Additional file 1: Figure S1) Fitzgerald Cat# 10R-G109A, RRID: AB 1285808 Goat polyclonal antiVEGF-C R & D systems Cat# AF752-SP Rabbit polyclonal antiRibosomal protein L 7a (E109) Cell signaling Technology Cat# 2415,RRID:AB_ 2182059 Rabbit polyclonal anti-eIF4G Gift from Dr. Rhoads (LSUHSC, Shreveport) Mouse monoclonal anti-eIF4A Rabbit polyclonal anti-Actin Sigma Cat# A2066, RRID: AB_476693 Chemicals Dulbecco’s modified eagle medium (DMEM) Sigma-Aldrich D6429 Antibiotic–Antimycotic (100X) ThermoFisher Scientific 15240-062
Techniques: Binding Assay, Western Blot, Control