fig3

Figure 3. (A) qRT-PCR analysis showing viral loads (PrM transcript levels) from SH-SY5Y recipient cells that were incubated (for three days) with infectious neuronal exosomes, pretreated overnight at 4°C and with different pH conditions (of 1.5, 4, 7, 9 or 11.5), respectively. All pH conditions were set in 1x PBS solution. Closed circles, squares, triangles, inverted triangles or rhombus represents viral loads obtained from LGTV-infected SH-SY5Y recipient cells incubated with infectious exosomes treated at indicated pH. In panel A, each data point represents data from one independent culture well in a plate. Each treatment had five independent replicates. The mRNA levels of prM gene are normalized to human beta-actin mRNA levels. P values less than 0.05 or equal to the numbers shown in the graph are from Student’s t-test. There is no statistical difference between groups of pH 4 and 9. Immunoblotting analysis is shown for proteins NS1 (B) or CD9 (C) and for all tested pH conditions. Asterisk (*) indicates glycosylated NS1 protein. Protein sizes are indicated in kilodaltons (kDa). Protein profile gel images in (B) and (C) are shown as loading controls.