Fig. 6: The modulatory role of Dll4 silencing on Notch signaling-mediated VSMC senescence through the suppression of palmitic acid-induced macrophage TLR4 pathway activation. | Nature Communications

Fig. 6: The modulatory role of Dll4 silencing on Notch signaling-mediated VSMC senescence through the suppression of palmitic acid-induced macrophage TLR4 pathway activation.

From: Palmitic acid in type 2 diabetes mellitus promotes atherosclerotic plaque vulnerability via macrophage Dll4 signaling

Fig. 6

A Representative images of immunofluorescent staining of CD68 and Dll4 in macrophages, with quantified expression of CD68 (left to right ***P < 0.001, ***P < 0.001, ***P < 0.001) and Dll4 (left to right ***P < 0.001, ***P < 0.001, ***P < 0.001, scale bar = 50 μm). B Western blots images and quantifications of relative levels of TLR4 (left to right *P = 0.0202, *P = 0.0298, P = 0.9882), FOXC2 (n left to right ***P < 0.001, ***P < 0.001, ns P = 0.9347), and Dll4 (left to right ***P < 0.001, ***P < 0.001, ***P < 0.001), as well as the relative phosphorylation level of ERK (left to right ***P < 0.001, ***P < 0.001, ns P = 0.8364). C A schematic diagram illustrating contact and non-contact co-culturing models of macrophages and VSMCs. D Immunofluorescent images of Dll4 in macrophages and NICD1 in VSMCs. Accompanying quantifications illustrate the relative Dll4 expression (left to right ***P < 0.001, ***P < 0.001), and the nuclear translocation of NICD1 (ns P = 0.30945, ***P < 0.001) in VSMCs, scale bar = 50 μm. E Western blots images and quantifications of relative expressions of HES1 (left to right ***P < 0.001, ***P < 0.001, ***P < 0.001), SIRT1 (left to right ***P < 0.001, ***P < 0.001, ***P < 0.001), P21 (left to right ***P < 0.001, ***P < 0.001, ***P = 0.0003), and the nuclear translocation of NICD1 (left to right ***P < 0.001, ***P < 0.001, **P = 0.0032) in VSMCs. F Immunofluorescent staining of NICD1 (scale bar = 50μm), SA-β-gal staining (scale bar = 50 μm), and CellEvent Green staining (scale bar = 40 μm) in VSMCs. The corresponding quantifications stand for nuclear translocation of NICD1 by co-localization analysis with DAPI in VSMCs (left to right ***P < 0.001, ***P < 0.001, ***P < 0.001), the extent of VSMC senescence by quantifying the percentage of SA-β-gal positive stained VSMCs (left to right ***P < 0.001, ***P < 0.001, *P = 0.0108) and mean fluorescent intensities of CellEvent Green staining (left to right ***P < 0.001, ***P < 0.001, ***P < 0.001). n = 6 independent replicates in all experiments, All data are presented as mean ± SD. A, B, E, F: one-way ANOVA with the Tukey post hoc correction, D: Two-tailed unpaired Student t-test and Two-tailed unpaired Student t-test with Welch’s correction. Source data are provided as a Source Data file.

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