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. 2022 Apr 28;18(4):e1010163. doi: 10.1371/journal.ppat.1010163

Fig 4. Colocalization and interaction of Loqs protein with DENV RNA.

Fig 4

(A) Fluorescent in situ hybridization imaging of Aag2 cells infected with DENV2 at an MOI of 1 after 48 hrs. NS3 and Loqs proteins (shown in green) were visualized using labeled antibodies, while DENV RNA (shown in red) was visualized using labeled antisense RNA probes. Costes p value was calculated to measure the extent of colocalization of DENV2 RNA with NS3/Loqs proteins. (B) Immunoprecipitation of HA-tagged Loqs from Aag2 cells infected with DENV2 at a MOI of 1. Aag2 cells transfected with HA-GFP or HA-Loqs PA/PB plasmids were infected with DENV2 for 48 hrs, and immunoprecipitations were performed with anti-HA antibodies. Abundances of HA-GFP and HA-Loqs in input lysates and immunoprecipitated material measured by western blot analysis. (C) DENV2 and RPL32 RNA abundances in immunoprecipitated RNA (IP) and input RNA (10%) were measured by semi-quantitative RT-PCR. A representative agarose gel image from three independent experiments is shown. (D) DENV2 RNA abundance in immunoprecipitated RNA (IP) as measured by RT-qPCR. Data was normalized to RPL32 mRNA levels (n = 3, ***p = 0.0006).