Journal: Experimental Biology and Medicine

Article Title: Engineering ADSCs by manipulating YAP for lymphedema treatment in a mouse tail model

doi: 10.3389/ebm.2024.10295

Figure Lengend Snippet: VEGFC successfully induced lymphatic endothelial transdifferentiation of ADSCs. (A–C) PCR tests showed the upregulation of VEGFR-3, Prox-1, Lyve-1 during the lymphatic endothelial transdifferentiation of ADSCs. (D) VEGFR-3 as a typical LEC marker was detected by immunofluorescence staining after VEGFC-induction at the indicated times. Scale bar = 50 μm. (E) Immunofluorescence intensity analysis of VEGFR-3. (F, H) Western blot showed the increased VEGFR-3 expression during the lymphatic endothelial transdifferentiation of ADSCs. (G) ADSCs were seeded on Matrigel after 7-day induction, and tube formation was evaluated at 12 h postseeding. VEGFC group generated tube-like structure while control group did not exhibit tubes. Scale bar = 100 μm. Bars: means ± standard deviation. n = 3 in each group, ns: no significant, ** P < 0.01, *** P < 0.001.

Article Snippet: For the lymphatic transdifferentiation of ADSCs, VEGFC (100 ng/mL, HY-P77864, MedChem Express, Monmouth Junction, NJ, United States) was used for 7 days.

Techniques: Marker, Immunofluorescence, Staining, Western Blot, Expressing, Generated, Control, Standard Deviation

Journal: Experimental Biology and Medicine

Article Title: Engineering ADSCs by manipulating YAP for lymphedema treatment in a mouse tail model

doi: 10.3389/ebm.2024.10295

Figure Lengend Snippet: Effect of lymphatic endothelial transdifferentiation and verteporfin on the expression of YAP. (A) Immunostaining of YAP in the control group and VEGFC group. Scale bar = 50 μm. (B) Immunofluorescence intensity analysis of YAP. (C–E) Western blot showed the decreased nuclear YAP expression and increased cytosolic YAP expression in ADSCs after lymphatic endothelial transdifferentiation. (F) PCR test showed that verteporfin suppressed the expression of YAP in ADSCs at the concentration of 20 μM. (G , H) Western blot showed the continuously inhibitory effect of verteporfin on YAP expression in ADSCs. Bars: means ± standard deviation. n = 3 in each group; ns, no significant, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: For the lymphatic transdifferentiation of ADSCs, VEGFC (100 ng/mL, HY-P77864, MedChem Express, Monmouth Junction, NJ, United States) was used for 7 days.

Techniques: Expressing, Immunostaining, Control, Immunofluorescence, Western Blot, Concentration Assay, Standard Deviation

Journal: Experimental Biology and Medicine

Article Title: Engineering ADSCs by manipulating YAP for lymphedema treatment in a mouse tail model

doi: 10.3389/ebm.2024.10295

Figure Lengend Snippet: The downregulation of YAP enhanced the lymphatic endothelial transdifferentiation of ADSCs in vitro . 20 μM verteporfin preconditioning for 48 h downregulated the expression of YAP in ADSCs. Under this inhibitory effect, higher expression levels of VEGFR-3 can be detected after differentiation of ADSCs, and larger density of tube formation can be observed. (A) Lymphatic endothelial transdifferentiation of ADSCs was conducted after 20 μM-verteporfin preconditioning for 48 h, and immunofluorescence staining indicated higher level of VEGFR-3. Scale bar = 50 μm. (B) Immunofluorescence intensity analysis of VEGFR-3. (C, D) Higher expression level of VEGFR-3 was confirmed by western blot. (E, F) The tube formation assay showed that VEGFC (+) verteporfin (+) group generated more tube-like structure than VEGFC (+) verteporfin (−) group. And quantification of master segments was analysed. Scale bar = 200 μm. Bars: means ± standard deviation. n = 3 in each group, ns: no significant, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: For the lymphatic transdifferentiation of ADSCs, VEGFC (100 ng/mL, HY-P77864, MedChem Express, Monmouth Junction, NJ, United States) was used for 7 days.

Techniques: In Vitro, Expressing, Immunofluorescence, Staining, Western Blot, Tube Formation Assay, Generated, Standard Deviation

Journal: Experimental Biology and Medicine

Article Title: Engineering ADSCs by manipulating YAP for lymphedema treatment in a mouse tail model

doi: 10.3389/ebm.2024.10295

Figure Lengend Snippet: YAP-downregulation ADSCs reduced the degree of swelling and improved fibrosis mouse tail lymphedema models. (A) Representative images of the mouse tail at 1, 2, 3, and 4 weeks after surgery. (B, C) Tail diameter was measured before and after surgery at the site of 5 mm and 10 mm distal from the incision. (D) Masson staining of the subcutaneous tissue of mouse tail at 2 and 4 weeks after surgery. Scale bar = 500 μm. (E) Statistical analysis of subcutaneous fibrosis ratio. (F) Representative results for Picro-Sirius red staining of mouse tail at 2 and 4 weeks after surgery. Red-yellow fibers represented collagen type I and green fibers with weak birefringence represented collagen type III. Scale bar = 20 μm. (G) Collagen type I/III ratio showed that more Collagen type III was produced in ADSCs (VEGFC+ verteporfin) group. n = 5 in each group, ns: no significant, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: For the lymphatic transdifferentiation of ADSCs, VEGFC (100 ng/mL, HY-P77864, MedChem Express, Monmouth Junction, NJ, United States) was used for 7 days.

Techniques: Staining, Produced