p38b and JAK-STAT signaling protect against Invertebrate iridescent virus 6 infection in Drosophila
p38-dependent IIV-6 induced unpaired expression A) S2* cells were infected with IIV-6 for the indicated times and induction of upd1, upd2, or upd3 was monitored by qRT-PCR, compared to PBS injected controls. Three biologically independent replicates are shown and statistical analysis was performed by Multiple t-tests with the Holm-Sidak correction for multiple comparisons. Error bars indicate standard deviation and black bars indicate the mean. B) TotA or TotM expression was measured by qRT-PCR from control w1118 flies as well as outstretcheds (oss), upd2Δ, upd3Δ, or upd2Δ,3Δ mutant flies 24 hours after IIV-6 infection or PBS injection. The results of 3–5 biologically independent assays are displayed. Error bars represent the standard deviation and black bars represent the mean. Statistical significance compared to the IIV-6 infected control strain (w1118) was determined by two-way ANOVA with Holm-Sidak correction. C) S2* cells were treated with inhibitors for the three MAPKs: JNK (SP600125, 25μΜ), ERK (U0125, 10μΜ), and p38 (SB203580, 10μΜ), or treated with a vehicle control (DMSO), for one hour prior to IIV-6 infection or mock treatment. After 24 hours of infection, upd3 expression was assayed by qRT-PCR. The results of 3 biologically independent assays are shown. Statistical significance, compared to the DMSO treated control, was determined by two-way ANOVA with Holm-Sidak correction. D) S2* cells were treated with the JAK inhibitor Tofacitinib (CP690,550 10 μM), or treated with a vehicle control (DMSO), for one hour prior to IIV-6 infection or mock treatment. After 24 hours of infection, TotA and upd3 expression were assayed by qRT-PCR. 3 biologically independent assays with statistical significance, comparing virus infected vehicle to inhibitor treatments, determined by two-way ANOVA with Holm-Sidak's multiple comparisons test. Significance is indicated in A-D as * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; and ns, not significant. A.U., Arbitrary Units.