Journal: PLOS Biology

Article Title: Translational repression of viral RNAs supports persistent arbovirus infection in mosquitoes

doi: 10.1371/journal.pbio.3003702

Figure Lengend Snippet: (A) Schematic representation of CHIKV RNA genome organization and viral protein expression strategy. Created in BioRender. Diez, J. (2026) https://BioRender.com/8srpphg . (B) CHIKV titers over time in U4.4 and C6/36, determined by plaque assay. (C) Kinetics of CHIKV gRNA and sgRNA levels in U4.4 and C6/36, quantified by RT-qPCR. (D and E) Expression of CHIKV nsP1 and capsid ( C ) proteins in U4.4 (D) and C6/36 (E) cells over the course of infection. Green and blue colors correspond to U4.4 and C6/36 cells, respectively. All infections were performed at an MOI of 4 and samples were collected at the indicated time post infection. Data points represent the mean ± SEM of 3 independent replicates. Viral RNA levels were quantified using a standard curve generated from in vitro -transcribed CHIKV RNA. Protein expression levels correspond to band intensities normalized to tubulin (Tub) and to the 1 dpi value within each blot. Dotted lines indicate separated lanes in the same membrane. All underlying data can be found in and in .

Article Snippet: The pellet was resuspended in RNAse-free water and consecutively treated with 0.1 μl Turbo DNase/μg RNA (ThermoScientific, AM1907) for 30 min at 37o C. Eighteen nanograms (ng) of total RNA was analyzed by TaqMan RT-qPCR using qScript XLT One-Step RT-qPCR ToughMix (Quanta BioSciences, 733-2232).

Techniques: Expressing, Plaque Assay, Quantitative RT-PCR, Infection, Generated, In Vitro, Membrane

Journal: PLOS Biology

Article Title: Translational repression of viral RNAs supports persistent arbovirus infection in mosquitoes

doi: 10.1371/journal.pbio.3003702

Figure Lengend Snippet: (A and B) Representative polysome profile analyses of U4.4 cells (A) and of C6/36 cells (B) at the indicated time points after CHIKV infection. (C and D) Polysome-to-monosome (P/M) ratios reflecting global mRNA translation in U4.4 and C6/36 cells over the course of infection calculated from the area under-the-curve (AUC) of polysome profiles and normalized to noninfected controls and to 1 dpi. (E) Levels of phosphorylated eIF2α in U4.4 and C6/36 cells during CHIKV infection were assessed by western blotting. Protein expression levels were normalized to tubulin (Tub) and to the noninfected value within each blot. Dotted lines indicate separated lanes in the same membrane. (F) P/M ratios of viral CHIKV gRNA and sgRNA in U4.4 and C6/36 cells over the course of infection were determined by RT-qPCR of RNA extracted from polysome fractions. Values are expressed relative to 1 dpi. Bars represent the mean ± SEM of 3 independent replicates. All underlying data can be found in and in .

Article Snippet: The pellet was resuspended in RNAse-free water and consecutively treated with 0.1 μl Turbo DNase/μg RNA (ThermoScientific, AM1907) for 30 min at 37o C. Eighteen nanograms (ng) of total RNA was analyzed by TaqMan RT-qPCR using qScript XLT One-Step RT-qPCR ToughMix (Quanta BioSciences, 733-2232).

Techniques: Infection, Western Blot, Expressing, Membrane, Quantitative RT-PCR