Fig.6. CHIP assay
Vero cells, in triplicates, were infected with BPXV at an MOI of 5,
followed by washing with PBS and addition of serum-free DMEM
supplemented with hesperetin or vehicle control. At 16 hpi, cell lysates
were prepared as described in method section and incubated with α-eIF4E
(reactive antibody), α- MNK1 (nonreactive antibody) or equivalent volume
of IP buffer (Beads control). This was followed by incubation with a
Protein A Sepharose® slurry. The beads were washed with IP buffer, and
cross-linking was reversed using Proteinase K. The reaction mixture was
centrifuged and the resulting supernatant was subjected to RNA
isolation, cDNA preparation, and quantitation of BPXV M gene by
qRT-PCR. Values are representative of three independent separate
experiments. Error bars indicate SD. Pair-wise statistical comparisons
were performed using the Student’s t test (*** = P<0.001).