Journal: Molecular Therapy. Nucleic Acids

Article Title: The miR-148/152 family contributes to angiogenesis of human pluripotent stem cell- derived endothelial cells by inhibiting MEOX2

doi: 10.1016/j.omtn.2023.04.020

Figure Lengend Snippet: Deletion of miR-148/152 family in ECs inhibits angiogenesis (A) Representative bright-field images of WT and TKO hESC-derived ECs at passage 4. (B) Total number of ECs at passages 1 to 4 (n = 4 per group). (C) EdU cell proliferation assays in WT and TKO hESC-derived ECs by flow cytometry. (D) Statistical analysis of EdU-positive ECs in EdU cell proliferation assays (n = 5 per group). (E) Representative bright-field images of wound-healing assay performed in WT and TKO hESC-derived ECs. (F) Quantitative statistics of migration area in the wound-healing assays (n = 3 per group). (G) Representative bright-field images of tube formation on Matrigel performed in WT and TKO hESC-derived ECs. (H) Statistical analysis of branch points formed by WT and TKO hESC-derived ECs (n = 3 per group). (I) Statistical analysis of the total length of capillary tubes formed by WT and TKO hESC-derived ECs (n = 3 per group). Scale bar indicates 200 μm. The data are expressed as the mean ± SEM. ∗∗∗p < 0.001.

Article Snippet: ECs were seeded on a solid layer of growth factor reduced Matrigel (Corning, USA) in a 96-well plate (1 × 10 4 cells per well), and cultured in ECM.

Techniques: Derivative Assay, Flow Cytometry, Wound Healing Assay, Migration

Journal: Molecular Therapy. Nucleic Acids

Article Title: The miR-148/152 family contributes to angiogenesis of human pluripotent stem cell- derived endothelial cells by inhibiting MEOX2

doi: 10.1016/j.omtn.2023.04.020

Figure Lengend Snippet: miR-152 overexpression restored the angiogenesis capacity of TKO hESC-derived ECs (A) Verification of miR-152 overexpression in TKO hESC-derived ECs by RT-qPCR (n = 5 per group). (B) Flow cytometric assay of EdU incorporation into TKO hESC-derived ECs after miR-152 overexpression. (C) Statistical analysis of EdU-positive ECs (n = 5 per group). (D) Representative bright-field images of wound-healing assay performed in TKO hESC-derived ECs with/without miR-152 overexpression. (E) Quantitative statistics of migration area in the wound-healing assays (n = 3 per group). (F) Representative bright-field images of tube formation on Matrigel performed in TKO hESC-derived ECs with/without miR-152 overexpression. (G) Statistical analysis of branch points formed in the tube formation assays (n = 3 per group). (H) Statistical analysis of the total length of capillary tubes formed in the tube formation assays (n = 3 per group). Scale bar indicates 200 μm. The data are expressed as the mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: ECs were seeded on a solid layer of growth factor reduced Matrigel (Corning, USA) in a 96-well plate (1 × 10 4 cells per well), and cultured in ECM.

Techniques: Over Expression, Derivative Assay, Quantitative RT-PCR, Flow Cytometry, Wound Healing Assay, Migration

Journal: Molecular Therapy. Nucleic Acids

Article Title: The miR-148/152 family contributes to angiogenesis of human pluripotent stem cell- derived endothelial cells by inhibiting MEOX2

doi: 10.1016/j.omtn.2023.04.020

Figure Lengend Snippet: MEOX2 suppression restores the angiogenesis capacity of TKO hESC-derived ECs (A) Verification of MEOX2 suppression in TKO hESC-derived ECs by RT-qPCR (n = 3 per group). (B) RT-qPCR detection of the downstream genes of MEOX2 in TKO hESC-derived ECs with/without MEOX2 suppression. (C) Flow cytometric assay of EdU incorporation into TKO hESC-derived ECs after transfection with MEOX2 shRNAs. (D) Statistical analysis of EdU-positive ECs in (C) (n = 5 per group). (E) Representative bright-field images of wound-healing assay performed in TKO hESC-derived ECs with/without MEOX2 shRNA transfection. (F) Quantitative statistics of migration area in the wound-healing assays (n = 5 per group). (G) Representative bright-field images of tube formation on Matrigel performed in TKO hESC-derived ECs transfected with/without MEOX2 shRNAs. (H) Statistical analysis of branch points formed in the tube formation assays (n = 3 per group). (I) Statistical analysis of the total length of capillary tubes formed in the tube formation assays (n = 3 per group). Scale bar indicates 200 μm. The data are expressed as the mean ± SEM. ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: ECs were seeded on a solid layer of growth factor reduced Matrigel (Corning, USA) in a 96-well plate (1 × 10 4 cells per well), and cultured in ECM.

Techniques: Derivative Assay, Quantitative RT-PCR, Flow Cytometry, Transfection, Wound Healing Assay, shRNA, Migration

Journal: Molecular Therapy. Nucleic Acids

Article Title: The miR-148/152 family contributes to angiogenesis of human pluripotent stem cell- derived endothelial cells by inhibiting MEOX2

doi: 10.1016/j.omtn.2023.04.020

Figure Lengend Snippet: The miR148/152 family regulates angiogenic capacity of hESC-derived ECs in vivo (A) Representative images of in situ (highlighted in circle) Matrigel plug at day 10 post injection of Matrigel mixed with WT, TKO, or miR-152-overexpressed hESC-ECs (n = 4). (B) Representative images of Matrigel plugs isolated from mice at day 10 post injection. (C) Representative H&E staining images of the explanted Matrigel plug sections. (D) Representative images of immunofluorescence staining for VWF (red) on the explanted Matrigel plug sections. Nuclei were stained with Hoechst 33342 (blue). Scale bar indicates 200 μm.

Article Snippet: ECs were seeded on a solid layer of growth factor reduced Matrigel (Corning, USA) in a 96-well plate (1 × 10 4 cells per well), and cultured in ECM.

Techniques: Derivative Assay, In Vivo, In Situ, Injection, Isolation, Staining, Immunofluorescence