Figure 3.

Intracellular complementation of Etf-1 PNA-reduced ehrlichial infection by ectopic expression of Etf-1-GFP. HEK293 cells expressing Etf-1 complemented the Etf-1 PNA knockdown and rescued E. chaffeensis infection. RNA samples prepared from HEK293 cells expressing Etf-1-GFP or GFP incubated with Etf-1 or CTL PNA-transfected E. chaffeensis were harvested at 48 h pi and subjected to qRT-PCR analysis (A) and protein analysis (B,C). The values reflect bacterial 16S rRNA and total Etf-1 normalized against human GAPDH mRNA, relative to the amount determined for the Etf-1 PNA treatment of HEK293 cells expressing Etf-1-GFP. Data indicate the mean ± standard deviation from three independent experiments performed in triplicate. ^*Significantly different (P < 0.05; analysis of variance) (A). Western blot analysis using anti-P28, actin, and Etf-1. The values under the bands show relative ratios of band intensities vs. actin, with the ratios of those from GFP plasmid and/or CTL PNA set to 1. (B) Results are presented as means ± standard deviations from three independent experiments. The asterisk indicates a significant difference compared with CTLs by analysis of variance (P < 0.01).