Journal: Frontiers in Cell and Developmental Biology

Article Title: mTORC1-Mediated Angiogenesis is Required for the Development of Rosacea

doi: 10.3389/fcell.2021.751785

Figure Lengend Snippet: mTORC1 signaling regulates LL37-induced cutaneous angiogenesis in vivo . (A,B) Immunostaining of pS6 and CD31 (A) , and quantitative analysis of pS6 and CD31 double positive cells (indicated by white arrows) (B) in skin tissues from healthy skin and rosacea patients. (C,D) Immunostaining of pS6 and CD31, and quantitation of pS6 and CD31 double positive cells (indicated by white arrows) in mice skin tissues treated with LL37 and/or RAPA. (E,F) Immunostaining of CD31, and quantitative analysis of the number of CD31 + vessels in control and LL37-treated rosacea-like mouse model treated with or without RAPA. (G,H) Immunostaining of CD31, and quantitation of CD31 + vessels in WT and TSC2 +/− mouse skin treated with or without LL37. (I) Vegf mRNA expression in WT and TSC2 +/− mouse skin treated with or without LL37 by RT-qPCR. (A,C,E) Right panels indicate the magnified picture in the left box with the same color. Scale bar: 50 μm. Data represent mean ± SEM from three independent experiments. Two-tailed unpaired Student’s t-test (B) or 1-way ANOVA with Bonferroni’s post hoc test (D,F,H,I) was used * p < 0.05 and ** p < 0.01.

Article Snippet: The membrane was blocked with 5% skimmed milk and then incubated with primary antibodies pS6 (5364, CST, 1:1000) and S6 (2217, CST, 1:1000) overnight with Tubulin (sc-5286, Santa Cruz Biotechnology, 1:1000) taken as a loading control.

Techniques: In Vivo, Immunostaining, Quantitation Assay, Control, Expressing, Quantitative RT-PCR, Two Tailed Test

Journal: Frontiers in Cell and Developmental Biology

Article Title: mTORC1-Mediated Angiogenesis is Required for the Development of Rosacea

doi: 10.3389/fcell.2021.751785

Figure Lengend Snippet: mTORC1 signaling regulates the tube formation mediated by LL37 in HUVECs. (A) HUVECs tube formation assay in presence of LL37. VEGF-treated group was set as a positive control. (B) Quantitative analysis of the tubular structure in (A) . (C) pS6 and S6 protein levels in LL37-treated HUVECs by western blotting. Tubulin was taken as a loading control. pS6 protein level was analyzed relative to total S6. (D) pS6 and S6 protein levels in LL37 and/or RAPA-treated HUVECs. Tubulin was taken as a loading control. pS6 protein level was analyzed relative to total S6. (E) HUVECs tube formation assay treated with LL37 and/or RAPA. (F) Quantitative analysis of the tubular structure in (E) . Scale bar: 50 μm. Data represent mean ± SEM from three independent experiments. Statistical significance was determined by One-way ANOVA (B,F) . * p < 0.05 and ** p < 0.01.

Article Snippet: The membrane was blocked with 5% skimmed milk and then incubated with primary antibodies pS6 (5364, CST, 1:1000) and S6 (2217, CST, 1:1000) overnight with Tubulin (sc-5286, Santa Cruz Biotechnology, 1:1000) taken as a loading control.

Techniques: Tube Formation Assay, Positive Control, Western Blot, Control