Skip to main content
. Author manuscript; available in PMC: 2020 May 14.
Published in final edited form as: Circulation. 2019 May 14;139(20):2326–2338. doi: 10.1161/CIRCULATIONAHA.118.035966

Figure 4. Analysis of early and late replication activities of 5’ terminally deleted, full-length, and mixed CV-B3/28 RNA populations in primary human cardiac cells.

Figure 4.

A–C. Synthetic RNAs (100 ng) of full-length (red squares) or deleted of 50 5’ terminal nucleotides (black circles) of CV-B3/28 forms were transfected alone or in association (95% of d50 associated with 5% of FL, blue triangles) in primary human cardiomyocytes (HCM): A. Viral RNA genomic replication activity (total viral RNA (intra and extracellular production)) was assessed using a RT-qPCR assay at 0, 8, 12, 24 and 48 hr post transfection. Data represent the mean ± SEM. *: P< 0.05 by Mann Whitney U test (n=3). B. Quantification by RT-qPCR of total, minus and EV RNA plus and minus strand ratios at each time post-transfection (in hours). RNA+/RNA− ratio values are indicated as an index above each sample. Data represent the mean ± SEM. Statistical analysis were assessed on the RNA+/RNA− ratios, *: P< 0.05; **: P< 0.01 by Mann Whitney U test (n=3). C. Viral RNA replication inhibition assay using guanidine hydrochloride (GuHCl; 2 mM) and fluoxetine (F; 8 μM) was performed at 0, 2 and 8 hr (h) post transfection for each viral form (d50: empty circle, FL: empty square, and mixed d50 + FL: empty triangle). Data represent the mean ± SEM. *: P< 0.05; **: P< 0.01; according to repeated measures two-way ANOVA test. D. Analysis of intra- and extra-cellular infectious particle production at 0, 8, 12, 24 and 48 hr (h) post transfection (n=3). Data represent the mean ± SEM. *: P< 0.05 by Mann Whitney U test.