Sequence-Specific Targeting of Bacterial Resistance Genes Increases Antibiotic Efficacy
(A) AcrA expression in E. coli with increasing concentrations of acrA-PPMO was quantified using an anti-AcrA antibody (top panel). AcrA expression was normalized against the expression of cAMP receptor protein (CRP). Error bars represent the standard deviations of normalized AcrA protein levels for six experimental replicates (middle panel). E. coli growth in fixed concentrations of clindamycin with increasing concentrations of acrA-PPMO is calculated by calculating the AUC growth in different experimental conditions (bottom panel). Error bars represent the standard deviation of growth rate changes of four experimental replicates. (B) acrA-PPMO has nonsignificant levels of toxicity to HBEC3KT human cells. HBEC3KT human cells were incubated with increasing doses of acrA-PPMO, and the number of viable cells was determined (Cell-Titer-Glo, Promega) every 24 h for 4 d. Error bars represent the standard deviation of cell counts obtained from ten replicate experiments.