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. 2019 Jun 12;10:642. doi: 10.3389/fphar.2019.00642

Figure 9.

Figure 9

Caspase-8 and annexinV activation in ZIKV-infected HUVECs. ZIKV replication was analyzed using Western blot (panel A) and qPCR (panel B). (A) Western blot analysis of envelop protein, caspase-8, and annexinV in ZIKV-infected HUVECs. HUVECs were infected with PRVABC59 and IBH30656 ZIKV. At 72 hpi, total proteins were collected and used for the detection of envelop protein, caspase-8, and annexinV. GAPDH was detected as the loading control. Lane 1: mock-infected HUVECs, lane 2: HUVECs infected with ZIKV strain PRVABC59; lane 3: HUVECs infected with ZIKV strain IBH30656. (B) qPCR analysis of virus transcript accumulation and caspase-8 transcription activation in ZIKV-infected HUVECs. Relative copies of each transcript were calculated using ∆∆Ct method. (C) IFA analysis of annexinV expression in ZIKV-infected HUVECs. A: mock-infected control; B: ZIKV strain PRVABC59; C: ZIKV strain IBH30656. Bar represents 10 µm size. (D) Effect of ZIKV replication on caspase-8 expression. UV-inactivated and replication-competent ZIKV PRVABC59 and IBH30656 were used to infect HUVECs. Total RNA was collected at 72 hpi and analyzed using qPCR. Relative copies of the viral genomes (ZIKV) were calculated using ∆∆Ct method. (E) Effect of ZIKV infection on cell vitality: mock-infected and ZIKV-infected HUVECs (IBH30656 and PRVABC59 strains) were counted using trypan blue. Experiments were performed in triplicate for three independent times.