Figure 4
The PERK 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 cells were lysed using TNET lysis buffer. Lysed samples were run on 12% SDS PAGE followed by Western blotting. Antibodies against phospho (Thr 980) PERK, phospho (Ser 51) eIF2α, eIF2α and CHOP were used to probe the PERK pathway component protein levels during CHIKV/SINV infection. Anti-actin antibody was used to probe loading control and uninfected cells (0h) were used as baseline protein level control. C, D) Under similar experimental conditions and time points stated above, real time RT PCR analysis of eIF2αK, CHOP and GADD34 transcripts was done on total RNA extracted from CHIKV/SINV infected cells using specific primers (Materials and Methods) against each of three genes. All three 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 at-least three independent repeats. Any significant change over 0h was determined using student unpaired T test and considered significant (*) if p<0.05.