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. 2021 Sep 29;9:693410. doi: 10.3389/fcell.2021.693410

FIGURE 4.

FIGURE 4

TWIST1-SLIT2 signaling mediates obesity-dependent inhibition of vascular formation in the gel subcutaneously implanted on mice. (A) 3D reconstructed immunofluorescence (IF) images showing GFP-labeled vascular formation and DAPI in the fibrin gel supplemented with GFP-labeled lean (BMI < 30, clone #15) vs. obese (BMI > 30, clone #7) human adipose ECs and subcutaneously implanted on NSG mice for 7 days. Scale bar, 50 μm. Graphs showing vascular area and average vessel length in the gel (n = 5–6, mean ± SEM, *p < 0.05). (B) 3D reconstructed IF micrographs showing GFP-labeled vascular formation and DAPI in the fibrin gel supplemented with GFP-labeled lean human adipose ECs (clone #15) treated with lentivirus encoding SLIT2 shRNA and subcutaneously implanted on NSG mice for 7 days. As a control, lean human adipose ECs were treated with control virus. Scale bar, 50 μm. Graphs showing vascular area and average vessel length in the gel (n = 6–7, mean ± SEM, *p < 0.05). (C) 3D reconstructed IF micrographs showing GFP-labeled vascular formation and DAPI in the fibrin gel supplemented with GFP-labeled obese human adipose ECs (clone #7) treated with SLIT2 protein, lentivirus overexpressing TWIST1 or in combination with SLIT2 shRNA and subcutaneously implanted on NSG mice for 7 days. As a control, obese human adipose ECs were treated with control virus (vector alone), control shRNA, or control vehicle. Scale bar, 50 μm. Graphs showing vascular area and average vessel length in the gel (n = 6–7, mean ± SEM, *p < 0.05).