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. Author manuscript; available in PMC: 2020 Jul 1.
Published in final edited form as: Ocul Surf. 2019 Mar 22;17(3):551–559. doi: 10.1016/j.jtos.2019.03.006

Figure 4. Effect of ISG15 in modulating ZIKV replication in Pr. HCECs.

Figure 4.

Pr. HCEC cells were transfected with scrambled or ISG15 siRNA followed by ZIKV infection (Brazilian strain BeH823339) at MOI 5. (A) Immunostaining for 4G2 antigen (green) was performed to detect ZIKV infectivity. (B) Quantification of ZIKV antigen (4G2) positive (+ve) cells per field upon ISG15 silencing. Silencing of ISG15 was confirmed by (C) qRT-PCR and (D&E) western blot followed by densitometry analysis. The values on bar graph (qPCR and densitometry) have been represented as mean ± SD; n=3, *** p<0.0005; * p<0.05. (F) Pr. HCEC cells were exogenously supplemented with rISG15 (100ng/ml) one hour prior to ZIKV infection. At 48h post infection, cells were stained using anti-flavivirus 4G2 antibody. (G) Quantification of average ZIKV antigen (4G2) positive (+ve) cells per field upon rISG15 supplementation (H) The culture supernatant was used for plaque assay and the values have been presented as mean ± SD; n=4, ** p<0.005; Student’s t-test.