Figure 3
The IRE-1 signaling during CHIKV and SINV infection. A, B) HEK293 cells (1×105 cells) were infected with MOI-1 of CHIKV/SINV and at indicated time points post infection total RNA was extracted to make the cDNA. Equal amounts (1μg each) of cDNA were used for PCR based XBP-1 splicing assay using specific primers against the spliced (s-XBP-1) and un-spliced (u-XBP-1) gene variants. Virus replications in the same samples (CHIKV/SINV) are probed using nsP1 specific primer for CHIKV or E1 specific primer for SINV (Materials and Methods). Actin gene amplification was used as an input RNA control and 0h (uninfected cells) was used as baseline control. C, D) Under similar experimental conditions and time points stated above, real time RT PCR analysis of XBP-1 and EDEM was done on total RNA extracted from CHIKV/SINV infected cells using specific primers (Materials and Methods) against each of the two genes. Both gene transcripts are presented as fold change over 0h (uninfected cells) after normalization with Actin and GAPDH mRNA. The graphs were plot using graph-pad prism software and are representative of three independent repeats. Any significant change over 0h was determined using student unpaired T test and considered significant (*) if p<0.05.