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. 2021 Aug 9;17(8):e1009729. doi: 10.1371/journal.pgen.1009729

Fig 2. Dll1 transactivates Notch which inhibits human myogenic differentiation.

Fig 2

(A) Schematics of heterologous cell-mixing assay and Notch signaling pathway. (B) DNA electrophoresis results that validated the specificity of human qPCR primers. (C) qPCR results of human HEY1 and HEYL using primers tested in B. Cells were differentiated for 24 hours after mixing. EV: empty control vector. The presence or absence of indicated cell types is indicated by plus or minus sign, respectively. n = 3. (D) Fluorescence images of mouse fibroblasts and human myoblasts in separate or mixing-culture conditions. Human myoblasts were labelled by red fluorescence protein Cherry; mouse fibroblasts (10T1/2) were infected by control (EV) or Dll1 expressing retroviruses. Cells were differentiated for 24 hours. Scale bar, 25 μm. (E) Quantification results of MyoG+ myoblasts as shown in D. n = 3. (F) Myosin immunostaining of human myoblasts that were differentiated for 96 hr. Scale bar, 100 μm. (G) Measurement of myoblast fusion as in F. n = 3. (H, I) qPCR results for genes in primary mouse myoblasts after mixing culture with fibroblasts. Data are means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001. ns, not significant.