AAV-mediated miR-199a inhibition in vivo
(A and B) Representative confocal images of wheat germ agglutinin (WGA, red) and DAPI (nuclei, blue) staining of hearts in 8-day-old, 4-week-old, and 13-week-old mice injected with AAV9-anti-miR-199a TuDs or AAV9-EGFP. Green, autofluorescence of cardiomyocytes with AAV9 virus infection. Scale bars, 200 μm; n = 4–5. (C) Calculated relative percentage of GFP-positive cells in the hearts of 8-day-old, 4-week-old, and 13-week-old mice injected with AAV9-anti-miR-199a TuDs or AAV9-EGFP. n = 4–5 different sections in each group. NS represents not significant compared with that in 8-day-old mice. (D) The expression of miR-199a in the hearts of 8-day-old, 4-week-old, and 13-week-old mice injected with AAV9-anti-miR-199a TuDs or AAV9-EGFP (n = 6). (E) Representative confocal images of WGA (red) and DAPI (nuclei, blue) staining of skeletal muscle, liver, and brain tissue in 13-week-old mice injected with AAV9-anti-miR-199a TuDs or AAV9-EGFP. Arrow, green autofluorescence of cardiomyocytes with AAV9 infection. Scale bars, 200 μm; n = 3. (F) The expression of miR-199a in liver, brain, and skeletal muscle of 13-week-old mice with AAV9-anti-miR-199a TuDs or AAV9-EGFP injection (n = 5). NS, not significant. (G) The plasma cytokine levels of IL-1β, IL-6, and IFN-γ in AAV-injected mice and wild-type control mice (n = 6). IFN-γ in each group was lower than 7.8 pg/mL. ∗p < 0.05, ∗∗p < 0.01.