Fig 6. ZIKV infection differentially alters certain astrocytic genes in U251 cells.

U251 cells were infected with ZIKV at MOI of 2 for indicated periods. (A) Cell viability was measured by the CCK-8 assay. Data represent as fold change compared to mock-treated cells. (B) ZIKV NS5 RNA was determined by qPCR with GAPDH as an internal control, and (C) viral dsRNA was immunoprobed with dsRNA-antibody (red), along with GFAP (green) for astrocytes and DAPI (blue) for nuclei by Immunofluorescent (IF), scale bar = 20 μm. (D and E) The mRNA expressions of PTBP1, GHR, LIF, PTBP3, EDNRB, and MBP were determined by qPCR (D) and protein expressions of PTBP1, GHR, LIF, EDNRB, and MBP was determined by Western blot analyses (E), respectively. In the qPCR analysis, the data represent the relative expression of target genes normalized to GAPDH as a reference internal control. In Western blot analysis, the β-actin was selected as an internal control. (F and G) Confocal microscopy images from MPAs stained with PTBP1, GHR, LIF, EDNRB, and MBP (green) for target genes and DAPI (blue) for nuclei, scale bar = 20 μm (F). Relative protein expression to control is quantified using Image J software (G). Data are shown as fold change from mock control. All the experiments were performed in triplicate, non-significant (ns); P < 0.05 (*); P < 0.01 (**); P < 0.001 (***). Student’s t-test. The underlying numerical data for “Fig 6A, 6B, 6D, and 6G” is provided in supporting file “S1 Data”.