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Figure 4. (A) qRT-PCR analysis showing viral loads (PrM transcript levels) from HaCaT recipient cells incubated with infectious tick exosomes treated at varying temperatures (of -80 °C, 4 °C, 12 °C, 23 °C, or 37 °C), and held overnight in 1 × chilled PBS with pH 7. All temperatures are represented as degree Celsius. Closed circles, squares, triangles, inverted triangles or rhombus represents viral loads obtained from LGTV-infected HaCaT cells incubated with infectious tick exosomes treated at indicated temperatures. 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 -80 °C and 4 °C, or 4 °C to 12 °C and 37 °C groups. Immunoblotting analysis is shown for proteins NS1 or CD63 (B) at all tested temperatures. Protein sizes are indicated in kilodaltons (kDa). Protein profile gel image in (B) is shown as the loading control.