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Figure 5 | Virology Journal

Figure 5

From: Differential unfolded protein response during Chikungunya and Sindbis virus infection: CHIKV nsP4 suppresses eIF2α phosphorylation

Figure 5

CHIKV infection suppresses the phosphorylation of eIF2α. A) Immunofluorescence microscopy at 10X magnification on CHIKV/SINV (MOI-1) or mock-infected HEK293 cells at 24 h. Unlike SINV, CHIKV infection failed to phosphorylate eIF2α (red). Virus infection was probed using dsRNA antibody (green). B) HEK293 cells were mock or CHIKV/SINV infected (MOI-1) till 12h to allow the translation of CHIKV encoded proteins followed by treatment with thapsigargin (0.1 μM) for 6h and Western blotting was performed on cell lysates using specific antibodies against phospho (Ser 51) eIF2α and eIF2α. Anti-actin antibody was used to probe loading control and uninfected or untreated cells (CC) were used as baseline control. Plaque assay titers in the presence of thapsigargin for 6h are presented as log pfu/ml. C) HEK293 cells were mock or CHIKV/SINV infected (MOI-1) for 1h. Cells were washed twice with 1X PBS to remove any traces of unbound virus particles followed by treatment with medium containing indicated concentrations of salubrinal for 24h. At 24 h media supernatant was used for plaque assay and cells were used for Western blotting using antibodies against phospho (Ser 51) eIF2α and eIF2α. Anti-actin antibody was used to probe loading control and uninfected or untreated cells (CC) were used as baseline control.

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