FIG 8.

Actin remodeling is important for HCMV-induced macropinocytosis. (A) HS-578T cells were pretreated with jasplakinolide (200 nM) for 60 min, followed by HCMV infection for 60 min. Virus internalization was then terminated by a low-pH buffer wash to inactivate any remaining extracellular virus. After an additional 5 h in culture, RNA was extracted and HCMV transcripts were detected using RT-qPCR and normalized against GAPDH amplified from the same reaction as described for Fig. 7. (B) The quantity of GAPDH RNA was determined by RT-qPCR to assess potential cytotoxicity. (C) HS-578T cells were pretreated with cytochalasin D (CytoD) at the indicated concentration, followed by HCMV infection for 4.5 h. RNA was extracted, and HCMV transcripts were detected using RT-qPCR and normalized against GAPDH amplified from the same reaction as an internal control. Infectivity was then normalized against that of the solvent control. (D) To assess potential cytotoxicity, the level of GAPDH RNA was determined by RT-qPCR at the highest CytoD concentration used for panel C (2.5 μM). (E) Cell viability was also monitored by CytoTox-One assay with 2.5 μM CytoD at the end of the infection.