%0 Figure %A de la Fuente, Cynthia %A Pinkham, Chelsea %A Dabbagh, Deemah %A Beitzel, Brett %A Garrison, Aura %A Palacios, Gustavo %A Hodge, Kimberley Alex %A Petricoin, Emanuel F. %A Schmaljohn, Connie %A Campbell, Catherine E. %A Narayanan, Aarthi %A Kehn-Hall, Kylene %D 2018 %T RPPA and Western blot confirmation of Smad protein phosphorylation. %U https://plos.figshare.com/articles/figure/RPPA_and_Western_blot_confirmation_of_Smad_protein_phosphorylation_/5859645 %R 10.1371/journal.pone.0191983.g002 %2 https://ndownloader.figshare.com/files/10412472 %K RVFV replication %K 913 genes %K Rift Valley fever virus infection Rift Valley fever virus %K decoy receptor %K hepatic fibrosis %K phosphorylation %K RPPA results %K interleukin 1 receptor type 2 %K host background %K TGF -β superfamily receptors %K RVFV infection %K Functional annotation %K gene promoter analysis %K phase protein array %K chromatin immunoprecipitation %K Smad protein family activation %K NS %K Smad-regulated genes %K Phosphoproteomic analysis %K IL 1R promoter %K attenuated RVFV strains %K axonal guidance %K Smad transcription factors %K cytoskeletal reorganization %K Smad proteins %K Smad-response element %X

A) Phosphorylation status of nine RPPA targets after RVFV infection of HSAECs. Bar graph represents the mean fold change with standard deviation of three independent replicates. Phospho-specific Smad antibodies are noted as: pSmad1/5/9 MH2 [phosphorylated Smad1/5 (S463/465)/9 (S465/467)], pSmad1/5 MH2 [phosphorylated Smad1/5 (S463/465)], pSmad2 MH2 [phosphorylated Smad2 (S465/467)], and pSmad2 L [phosphorylated Smad2 (S245/250/255)]. Statistical significance of the phosphorylation changes are highlighted in Table 1. HSAEC (B) and Huh-7 cells (C) were infected at an MOI 5 with MP12 or ZH548, or mock infected (media alone). Protein lysates harvested at 3, 9, and 18hpi were analyzed for pSmad1/5/9 MH2, pSmad1/5 MH2, pSmad2 MH2, pSmad2 L, RVFV NP, Actin as well as total protein levels for Smad1, Smad5, and Smad2 by western blotting. Blots represent one replicate of three independent experiments. M:Mock; MP:MP12; ZH: ZH548.

%I PLOS ONE