Figure 5

Bacteria-induced gene expressions of the putative clotting factors. Fifth instar nymphs were microinjected with E. coli K12 or B. subtilis. Total RNA was extracted from the nymphs at the indicated times after injection. PBS-injected sample was used as a control. The relative expression levels of each gene at different time points were normalized using the N. lugens 18 s rRNA Ct values. Three biological replications were conducted and the relative transcript levels at each time point were calculated using the ΔΔCt method. Results of triplicate experiments are shown with the standard deviations. The asterisk (**) indicates statistical significance at p < 0.01 between the PBS-injected and bacteria-injected nymphs. *indicates statistical significance at p < 0.05 between the PBS-injected and bacteria-injected nymphs. The E. coli K12 and B. subtilis injected samples are shown on the left (black) and right (dark gray), respectively. C refers to the PBS-injected control. 6, 12 and 24 h refer to bacteria-injected specimens at 6, 12 and 24 h p.i.