Journal: bioRxiv

Article Title: A global lipid map defines a network essential for Zika virus replication

doi: 10.1101/2020.01.27.910919

Figure Lengend Snippet: Sphingolipids are essential for ZIKV infection. a Overview of sphingolipid biosynthesis. SPT, serine palmitoyltransferase; KDSR, 3-ketodihydrosphingosine reductase; DegS, Delta 4-desaturase, Sphingolipid; CerS, ceramide synthase; SGMS1, sphingomyelin synthase 1; CERT, ceramide transfer protein. b , c Huh7 cells treated with myriocin, FB1, or a vehicle control were infected with ZIKV (MOI = 0.1). At the indicated times post-infection, culture supernatants were collected and analyzed by plaque assay ( c ) or RT-qPCR ( d ); n = 3 independent experiments. Two-way ANOVA with Dunnett’s multiple comparisons test. d Vero cells treated with myriocin, FB1, or vehicle were infected with ZIKV (MOI = 0.1). At 72 hpi, intracellular levels of ZIKV E protein were assessed by immunoblotting. Blot is representative of 2 independent experiments. e , f HAP1 human ( e ) and DC2.4 murine dendritic ( f ) WT and SPTLC2-knockout cells were infected with ZIKV (MOI = 0.1). At the indicated timepoints, culture supernatants were collected and analyzed by plaque assay; n = 3 independent experiments. Two-way ANOVA with Sidak’s multiple comparisons test (e) and two-tailed Student’s t -test (f). g iPSC-derived human neural progenitor cells (hNPCs) treated with myriocin, FB1 or vehicle were infected with ZIKV (MOI = 0.1). At the indicated times post-infection, culture supernatants were collected and analyzed by plaque assay; n = 3 independent experiments. Two-way ANOVA with Tukey’s multiple comparisons test. h SH-5YSY human neuroblastoma cells were treated with inhibitors, infected with ZIKV (MOI = 0.1) and analyzed by plaque assay. Two-way ANOVA with Tukey’s multiple comparisons test. Data are mean ± SD. See also Supplementary Data 3, Supplementary Fig. 4, Supplementary Fig. 5, and the Source Data file.

Article Snippet: Human iPSC (XCL-1) -derived neural progenitor cells (hNPCs) were purchased commercially (StemCell Technologies, Cat #70901, Lot #17080) and maintained up to ten passages according to the manufacturer’s instructions.

Techniques: Infection, Plaque Assay, Quantitative RT-PCR, Western Blot, Knock-Out, Two Tailed Test, Derivative Assay

Journal: Nature Communications

Article Title: A global lipid map defines a network essential for Zika virus replication

doi: 10.1038/s41467-020-17433-9

Figure Lengend Snippet: a Overview of sphingolipid biosynthesis. SPT serine palmitoyltransferase, KDSR 3-ketodihydrosphingosine reductase, DegS delta 4-desaturase, sphingolipid, CerS ceramide synthase, SGMS1 sphingomyelin synthase 1, CERT ceramide transfer protein. b , c Huh7 cells treated with myriocin, FB1, or a vehicle control were infected with ZIKV (MOI = 0.1). At the indicated times post infection, culture supernatants were collected and analyzed by plaque assay ( b ) or RT-qPCR ( c ); n = 3 independent experiments. Two-way ANOVA with Dunnett’s multiple-comparison test. d Vero cells treated with myriocin, FB1, or vehicle were infected with ZIKV (MOI = 0.1). At 72 hpi, intracellular levels of ZIKV E protein were assessed by immunoblotting. Blot is representative of two independent experiments. e , f HAP1 human ( e ) and DC2.4 murine dendritic ( f ) WT and SPTLC2-knockout cells were infected with ZIKV (MOI = 0.1). At the indicated timepoints, culture supernatants were collected and analyzed by plaque assay; n = 3 independent experiments. Two-way ANOVA with Sidak’s multiple-comparison test ( e ) and two-tailed Student’s t test ( f ). g iPSC-derived human neural progenitor cells (hNPCs) treated with myriocin, FB1, or vehicle were infected with ZIKV (MOI = 0.1). At the indicated times post infection, culture supernatants were collected and analyzed by plaque assay; n = 6 independent infections. Two-way ANOVA with Tukey’s multiple- comparison test. h SH-5YSY human neuroblastoma cells were treated with inhibitors, infected with ZIKV (MOI = 0.1), and analyzed by plaque assay. Two-way ANOVA with Tukey’s multiple-comparison test. Data are mean ± SD. See also Supplementary Data , Supplementary Fig. , Supplementary Fig. , and the Source Data file.

Article Snippet: Human iPSC (XCL-1) -derived neural progenitor cells (hNPCs) were purchased commercially (StemCell Technologies, Cat #70901, Lot #17080) and maintained up to ten passages according to the manufacturer’s instructions.

Techniques: Control, Infection, Plaque Assay, Quantitative RT-PCR, Comparison, Western Blot, Knock-Out, Two Tailed Test, Derivative Assay