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. Author manuscript; available in PMC: 2021 Mar 15.
Published in final edited form as: Science. 2020 Nov 20;370(6519):eabc3621. doi: 10.1126/science.abc3621

Figure 6. Purinergic signaling contributes to rotavirus disease processes.

Figure 6.

(A) Rotavirus yield from SA114F-infected MA104 cells treated with DMSO vehicle, 10U/mL apyrase, 10μM BPTU, 10μM AR-C 118925XX, 20μM Bx430, 20μM 5-BDBD, 300μM suramin, or 10μM PPADS by fluorescent focus assay (data combined from N=3 independent experiments). (B) Plaque assay yield of RV (SA114F)-infected MA104-GCaMP (Par) or MA104-GCaMP P2Y1 knockout (KO) cells (data combined from N=3 independent experiments). (C-E) qPCR of mRNA transcripts normalized to 18S mRNA transcripts and fold change relative to the Mock-DMSO transcript levels in (J3)HIE monolayers mock- or RV (Ito)-infected and treated with DMSO vehicle, 100U/mL apyrase, or 10 μM BPTU (data combined from N=3 independent experiments). (F-G) Serotonin secretion from RV (Ito)-infected jHIE (F) monolayers and (G) transwells treated with DMSO, 100U/mL apyrase, 10μM BPTU, or 300nM ω-agatoxin (data combined from N=3 independent experiments). (H) Enteroid swelling assay: 3D jHIE-GCaMP6s enteroids mock- or RV (Ito)-infected and treated with DMSO or 10μM BPTU. Cross-sectional area of the internal lumen (pink outline) used for percent increase between basal and max swelling (left panels). Scale bar = 100μm. (n ≥ 68 HIEs per condition, data combined from N=3 independent experiments) (I-K) C57Bl/6J mouse pups with diarrhea infected with Rhesus RV and vehicle- or BPTU-treated and the (J) mean diarrhea score. (data combined from 4 cages of each condition, 26–30 pups total per condition, mean ± SEM) (K) Summary model of RV-induced ICWs mediated by extracellular ADP. (A-B, F-G) One-way ANOVA with Bonferroni multiple comparisons test, (C-J) Kruskal-Wallis with Dunn’s multiple comparisons, or (I-J) Mann-Whitney tests used. (A-H) Data represented as mean ± SD, (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001).