Fig. 3: miR-200c-3p delivery to lung endothelial cells promotes ATII transdifferentiation through down-regulation of Flt1.

A Representative immunofluorescence images of control ATII cells (stained for Rage, red) co-cultured with ECs (stained for Erg, green) either untreated or transfected with the indicated miRNAs. B Quantification of fully differentiated (>1500 µm2) Rage+ ATI cells, expressed as a percentage of Rage+ cells, in the indicated conditions; n ≥ 9. C Representative immunofluorescence images of ATII cells from bleomycin-treated mice (stained for Rage, red) co-cultured with ECs (stained for Erg, green) either untreated or transfected with the indicated miRNAs; n ≥ 9. D Quantification of fully differentiated (>1500 µm2) Rage+ ATI cells, expressed as a percentage of Rage+ cells, in the indicated conditions; n ≥ 9. E Real-time PCR quantification of the expression levels of Flt1 in lungs harvested from control mice and mice treated with bleomycin, either in the presence or in the absence of miR-200c. Results are normalized to Gapdh expression; n = 4. F Representative immunofluorescence images of control ATII cells kept in culture with either wt or Flt1 KO EC, stained for Erg (green) and Rage (red), respectively. Nuclei were counterstained with Hoechst (blue). G Quantification of fully differentiated (>1500 µm2) Rage+ ATI cells, expressed as a percentage of Rage+ cells, in the indicated conditions; n = 9. H Representative immunofluorescence images of ATII cells from bleomycin-treated mice, kept in culture with either wt or Flt1 KO EC, stained for Erg (green) and Rage (red), respectively. Nuclei were counterstained with Hoechst (blue). I Quantification of fully differentiated (>1500 µm2) Rage+ cells ATI cells from bleomycin mice, expressed as a percentage of Rage+ cells, in the indicated conditions; n ≥ 9. Scale bar in A, C, F, H, 100 μm. Data in B, D, E, G, and I are shown as mean ± S.E.M. Statistical significance was determined using one-way ANOVA, ***P < 0.001, ****P < 0.0001.