MOESM1 of HSV-1 triggers paracrine fibroblast growth factor response from cortical brain cells via immediate-early protein ICP0
Creators: name: Hensel, Niko
name: Raker, Verena
name: Förthmann, Benjamin
name: Detering, Nora Tula
name: Kubinski, Sabrina
name: Buch, Anna
name: Katzilieris-Petras, Georgios
name: Spanier, Julia
name: Gudi, Viktoria
name: Wagenknecht, Sylvia
name: Kopfnagel, Verena
name: Werfel, Thomas
name: Stangel, Martin
name: Beineke, Andreas
name: Kalinke, Ulrich
name: Paludan, Søren
name: Sodeik, Beate
name: Claus, Peter
Publisher: name: figshare
Year: 2019
Subjects: subject: Biochemistry
subject: Medicine
subject: Microbiology
subject: FOS: Biological sciences
subject: FOS: Biological sciences
subject: Cell Biology
subject: Genetics
subject: Molecular Biology
subject: Neuroscience
subject: Immunology
subject: FOS: Clinical medicine
subject: FOS: Clinical medicine
subject: Developmental Biology
subject: 110309 Infectious Diseases
subject: FOS: Health sciences
subject: FOS: Health sciences
subject: 60506 Virology
subject: Computational Biology

description: Additional file 1: Figure S1. mRNAs of housekeeping genes and some FGFs become down-regulated in HSV-1 infected PCCs. A, Negatively regulated FGF mRNAs in HSV-1(17+)Lox (MOI 10) infected PCCs, mean ± SEM of technical duplicates. B, mRNAs of housekeeping genes GAPDH and PPIA in PCCs infected with HSV-1(17+)LOX. C, mRNA levels of GDNF and NGF in infected. Mean ± SEM, n=3, two-way ANOVA, Holm-Sidak‘s multiple comparison test (* p&lt;0.05, ** p&lt;0.01, *** p&lt;0.001). Figure S2. FGF-4 is correctly spliced and expressed in HSV-1 infected PCCs. HSV-1 randomly provokes a dysregulation of transcription termination producing non-translated transcripts including introns [79]. A, Two primer sets were used for amplification of spliced and non-spliced FGF-4 transcripts in qRT-PCR (black arrows) and conventional RT-PCR (red arrows). B, cDNA and RNA from control and HSV-1(17+)Lox infected PCCs (MOI 10, 6 hpi) was used with primers able to amplify intron sequences (red arrows in A). NTC = non template control. C, Fold change of FGF-4 mRNA in HSV-1(17+)Lox infected PCCs (MOI10). Mean ± SEM, one-way ANOVA, Holm-Sidak‘s multiple comparison test (****p&gt;0.0001). Figure S3. Filtration of HSV-1 conditioned medium removes viral particles. The media of HSV-1 infected cells were filtered followed by plaque assays. We could observe plaques in non-filtered media (detection limit 40 PFU/ml), but no plaques in cells incubated with filtered, HSV-1 conditioned medium. Mean ± SEM, n=5. Figure S4. Increasing doses of UV-radiation de-activate HSV-1. Transcript level of different viral genes in PCCs infected with untreated or UV-inactivated HSV-1(17+)Lox (MOI 10, 4 hpi). Mean from three pooled replicates. Figure S5. FGF-4 responses are not triggered by MyD88 dependent TLRs or cGAS/STING innate sensing mechanisms. A, PCCs were incubated with TLR agonists for 6 hours and FGF-4 mRNA levels were quantified. Mean, n=1. B, PCCs derived from STING or MyD88 knock-out mice infected with HSV-1 (MOI 10). Mean ± SEM, n=5, one-way ANOVA, Holm-Sidak‘s multiple comparison test (*** p &lt; 0.001, ** p &lt; 0.01, **** p &lt; 0.0001). Table S1. PCR-primer sequences for the detection of murine transcripts or HSV-1 transcripts.