Fig. 5: TET-dependent demethylation of fibroblast-borne endothelial genes in dermal fibroblast causes the formation of vasculogenic fibroblast and rescues perfusion in diabetic ischemic limbs.
Representative PeriMed laser speckle–assisted limb perfusion images at d14 (A) and its quantification (B) of hind limb perfusion at different time points post-surgery in db/db mice treated with TNTEP+scramble, TNTEFF + scramble, TNTEFF + Tet1/2/3 shRNA and TNTEP + Tet1/2/3 shRNA post hind-limb surgery (n = 7). *TNTEFF + scramble vs TNTEP+scramble; §TNTEFF + scramble vs TNTEFF + Tet1/2/3 shRN, #TNTEFF + scramble vs TNTEP + Tet1/2/3 shRNA (one-way ANOVA, Tukey HSD post-hoc-test). One-way ANOVA, followed by Tukey HSD post-hoc-test). The study design and time points are similar to Fig. 3A. C Representative ultrasound and flow velocity images and quantification (D) of the flow velocity of the hind limb in db/db mice treated with combinations mentioned in (B) (n = 7, 6, 7, 6). One-way ANOVA, Tukey HSD post-hoc-test. Results represent mean ± S.D. E Immunofluorescence images of hind-limb ischemic skin and colocalization quantification (F) of COL1A2 (green) and VWF (red) staining of the abovementioned groups (B) were determined by Pearson correlation coefficient (r). The white arrow represents the area of colocalization. Data expressed as mean ± S.D (n = 6, 6, 7, 6). One-way ANOVA, Tukey HSD post-hoc-test). AU, arbitrary unit. G Immunofluorescence confocal image of day 14 hind limb ischemic tissue in TNTEFF+scramble db/db mice stained for COL1A2 and VWF. The left panel shows the 3D-reconstruction image of the colocalized COL1A2 and VWF rendered by IMARIS software (inset). The middle panel shows the magnified inset. The right panel shows the colocalized area for COL1A2 and VWF. Scale, 20 µm. Source data are provided as a Source Data file.
