Heat shock protein 90ß in the Vero cell membrane binds Japanese encephalitis virus

Wang, Yuan; Li, Yan; Ding, Tianbing

Decreased HSP90β expression in Vero cells reduces JEV progeny yields. siRNAs subcloned in lentivirus vectors were implemented to reduce HSP90β expression in Vero cells, and one transfected cell line exhibited an over 50% steady inhibition of HSP90β expression (Fig. 4A) . Subsequent sequencing data revealed that the 21-nt siRNA (GGAUGACA GCGGUAAGGAUtt) was complementary to the human HSP90β mRNA 1012-1030 nt (GGATGACAGTGGTAA GGATAA), which is consistent with a previous report (31) . Overall, the progeny virions in both intact and siRNA-treated Vero cell culture supernatants were detected at 24 (Fig. 4B ), 36 and 48 hpi. The commensurate numbers of viral plaques at 36 and 48 hpi in the intact cells indicated that fewer infectious viruses were released, implicating that almost all cells became infected after 36 hpi when the MOI was 0.1. However, in the siRNA-treated cells, the virion yields were significantly inhibited (P<0.01), particularly at 24 and 36 hpi, by 6.5-and 5.6-fold, respectively (Fig. 4C ).