FIG 3.

YBX1 interacts with DENV vRNA and capsid protein but is dispensable for nucleocapsid formation. Huh7 WT and YBX1 KO cells were infected with DENV at MOI 5 and processed for RNA-immunoprecipitation (RIP) or Co-IPs at 24 hpi. Cell lysates were incubated with an isotype control (IgG) and an antibody against YBX1 or capsid. Bound material was captured on Dynabeads protein G. (A) YBX1-RIP. Representative immunoblot of YBX1-pull down and total number of vRNA copies as determined in the IP by RT-qPCR. (B) IgG and YBX1-IPs were left untreated or treated with RNase A/T1 and immunoblots were performed for YBX1, capsid and envelope. (C) Densitometry analysis was performed to determine the levels of YBX1 and capsid in the IP sample treated with RNase relative to the nontreated control (lane 7 versus 6). (D) Capsid-RIP. Representative immunoblot of capsid-pull down and total number of vRNA copies as determined in the IP by RT-qPCR in WT and YBX1 KO cells. Data are presented as mean ± SEM from three independent experiments.