ATCC vr 1238 emcv emc strain atcc

Figure 4. HIF activation exerts broad antiviral activity in neuron-like cells (A and B) Virus titer of vesicular stomatitis virus (VSV) and <t>encephalomyocarditis</t> <t>virus</t> <t>(EMCV)</t> in culture supernatants from SH-Sy5y cells treated with ML228 (0.5 mM) and infected for 19 h with EMCV (MOI 0.1) or VSV (MOI 0.001). n = 3 biological replicates. (C) SARS-CoV-2 N2 RNA levels evaluated by RT-qPCR in total RNA from SH-Sy5y cells treated with ML228 (0.5 mM) and infected for 48 h with SARS-CoV-2 (MOI 1). Data were normalized against 18S rRNA. n = 5 biological replicates. (D) Inﬂuenza A virus M2 RNA levels evaluated by RT-qPCR in total RNA from SH-Sy5y cells treated with ML228 (0.5 mM) and infected for 24 h with inﬂuenza A virus (MOI 0.1). Data were normalized against 18S rRNA. n = 5 biological replicates. (E) Measles virus titer in culture supernatants from SH-Sy5y cells treated with ML228 (0.5 mM) and infected for 36 h with Measles virus (MOI 0.1). n = 6 biological replicates. Data are representative of at least three independent experiments. The data are shown as means ± SD. Individual values are represented by dots. For (A), Mann- Whitney was applied to identify statistical signiﬁcance. For (B)–(E), the statistical analyses were performed by Student’s t test, two-tailed, parametric distribution to identify statistical signiﬁcance. ns, not signiﬁcant. Illustrations of viruses were generated with BioRender.

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Figure 4. HIF activation exerts broad antiviral activity in neuron-like cells (A and B) Virus titer of vesicular stomatitis virus (VSV) and encephalomyocarditis virus (EMCV) in culture supernatants from SH-Sy5y cells treated with ML228 (0.5 mM) and infected for 19 h with EMCV (MOI 0.1) or VSV (MOI 0.001). n = 3 biological replicates. (C) SARS-CoV-2 N2 RNA levels evaluated by RT-qPCR in total RNA from SH-Sy5y cells treated with ML228 (0.5 mM) and infected for 48 h with SARS-CoV-2 (MOI 1). Data were normalized against 18S rRNA. n = 5 biological replicates. (D) Inﬂuenza A virus M2 RNA levels evaluated by RT-qPCR in total RNA from SH-Sy5y cells treated with ML228 (0.5 mM) and infected for 24 h with inﬂuenza A virus (MOI 0.1). Data were normalized against 18S rRNA. n = 5 biological replicates. (E) Measles virus titer in culture supernatants from SH-Sy5y cells treated with ML228 (0.5 mM) and infected for 36 h with Measles virus (MOI 0.1). n = 6 biological replicates. Data are representative of at least three independent experiments. The data are shown as means ± SD. Individual values are represented by dots. For (A), Mann- Whitney was applied to identify statistical signiﬁcance. For (B)–(E), the statistical analyses were performed by Student’s t test, two-tailed, parametric distribution to identify statistical signiﬁcance. ns, not signiﬁcant. Illustrations of viruses were generated with BioRender.

Journal: Cell reports

Article Title: The HIF transcription network exerts innate antiviral activity in neurons and limits brain inflammation.

doi: 10.1016/j.celrep.2024.113792

Figure Lengend Snippet: Figure 4. HIF activation exerts broad antiviral activity in neuron-like cells (A and B) Virus titer of vesicular stomatitis virus (VSV) and encephalomyocarditis virus (EMCV) in culture supernatants from SH-Sy5y cells treated with ML228 (0.5 mM) and infected for 19 h with EMCV (MOI 0.1) or VSV (MOI 0.001). n = 3 biological replicates. (C) SARS-CoV-2 N2 RNA levels evaluated by RT-qPCR in total RNA from SH-Sy5y cells treated with ML228 (0.5 mM) and infected for 48 h with SARS-CoV-2 (MOI 1). Data were normalized against 18S rRNA. n = 5 biological replicates. (D) Inﬂuenza A virus M2 RNA levels evaluated by RT-qPCR in total RNA from SH-Sy5y cells treated with ML228 (0.5 mM) and infected for 24 h with inﬂuenza A virus (MOI 0.1). Data were normalized against 18S rRNA. n = 5 biological replicates. (E) Measles virus titer in culture supernatants from SH-Sy5y cells treated with ML228 (0.5 mM) and infected for 36 h with Measles virus (MOI 0.1). n = 6 biological replicates. Data are representative of at least three independent experiments. The data are shown as means ± SD. Individual values are represented by dots. For (A), Mann- Whitney was applied to identify statistical signiﬁcance. For (B)–(E), the statistical analyses were performed by Student’s t test, two-tailed, parametric distribution to identify statistical signiﬁcance. ns, not signiﬁcant. Illustrations of viruses were generated with BioRender.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies HIF-1alpha (D2U3T) Cell Signaling #14179S; RRID:AB_2622225 THOC1 Sigma-Aldrich #HPA019687-25UL; RRID:AB_1857980 Vinculin Sigma-Aldrich #V9131-0.2ML; RRID:AB_477629 LC3B Cell Signaling #2775; RRID:AB_915950 ATG5 (D5F5U) Cell Signaling #12994; RRID:AB_2630393 VHL Cell Signaling #68547; RRID:AB_2716279 BNIP3 Cell Signaling #44060; RRID:AB_2799259 STAT1 Cell Signaling #9172; RRID:AB_2198300 pSTAT1 Cell Signaling #7694S; RRID:AB_10950970 IFNAR2 Abcam #ab193410 TBK1 Cell Signaling #38066; RRID:AB_2827657 pTBK1 Cell Signaling #5483; RRID:AB_10693472 V5 Abcam #ab27671; RRID:AB_471093 IRF3 Cell Signaling #4302; RRID:AB_1904036 pIRF3 Cell Signaling #37829; RRID:AB_2799121 STING Cell Signaling #13647; RRID:AB_2732796 pSTING Cell Signaling #50907; RRID:AB_2827656 peroxidase-conjugated F(ab)2 donkey anti-mouse IgG (H + L) Jackson ImmunoResearch #PRID: AB_2340774 peroxidase-conjugated F(ab)2 donkey anti-rabbit IgG (H + L) Jackson ImmunoResearch #PRID: AB_2340590 LC3 Nordic Biosite #PM036; RRID:AB_2274121 P62 Progen #GP62-C; RRID:AB_2687531 ICP5/VP5 Abcam Ab6508 b-Tubulin Cell Signaling #2146; RRID:AB_2210545 Alexa 488 Thermo Fischer #A-21206; RRID:AB_2535792 Alexa A568 Thermo Fischer #A-11075; RRID:AB_2534119 DAPI Santa Cruz #sc-3598 goat anti-mouse IgG1 biotin SouthernBiotech #1071-08; RRID: AB_2794427 Bacterial and virus strains HSV-1 (KOS) strain ATCC # VR-1493 HSV-2 (333) strain Kristina Eriksson Iversen et al.6 Clinical isolates HSV-1 2762 Kristina Eriksson Iversen et al.6 HSV-2 B4327UR Kristina Eriksson Iversen et al.6 VSV (Indiana) strain ATCC #VR-1238 EMCV (EMC) strain ATCC #VR-129B Influenza virus A (H1N1), PR8 strain ATCC #VR-1469 Measles virus (Edmonston) strain ATCC #VR-24 SARS-CoV-2 (MZ314997/B.1.1.7 variant) Arvind Patel, University of Glasgow SARS-CoV-2 B1.1.7 Chemicals, peptides, and recombinant proteins CoCl2 Sigma-Aldrich #14726-62-6 Diethyl succinate Sigma-Aldrich #123-25-1 Dimethyl sulfoxide (DMSO) Sigma-Aldrich #67-68-5 Vadadustat MedChemExpress #HY-101277 (Continued on next page) Cell Reports 43, 113792, February 27, 2024 17

Techniques: Activation Assay, Activity Assay, Virus, Infection, Quantitative RT-PCR, MANN-WHITNEY, Two Tailed Test, Generated

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