FIG 8.

Salmonella engages members of the miR30 family to downregulate Ubc-9. (A) Quantitative real-time PCR analysis of mature miR30c and miR30e upon S. Typhimurium infection at different time points. SnoRNAs SNORD61 and RNU6 were used for normalization. A representative experiment of three replicates is depicted. (B) Immunoblot analysis of Ubc-9 was carried out in cells transfected with 30 nM concentrations of the mimics of miR30c and miR30e. GAPDH was used for normalization. Numbers at left of panels B and C are molecular masses in kilodaltons. (C) Ubc-9 levels were assessed in HCT-8 cells upon infection with S. Typhimurium with or without inhibitors of the miRNAs miR30c and miR30e. GAPDH was used for normalization. (D and E) GPA to assess bacterial load was performed after S. Typhimurium infection in HCT-8 cells transfected with mimics (D) or inhibitors (E) of miR30c and miR30e. CFU were plotted, and statistical analyses were carried out using Student's t test. The asterisk indicates a P value of ≤0.05, and the number sign indicates a P value of ≤0.01.