Fig 2: Knock-down of MFN2 in ApoE−/− mice aggravated vascular senescence by affecting the expression of P53 and P21(A) To investigate the effect of Ang II stimulation on the expression of BCL6, MFN2, P53, and P21 proteins in mice aorta and (B) quantification.(B) Effects of shMfn2 followed by Ang II stimulation on MFN2, P53, and P21 protein expression in mice aorta and (D) quantification. In (B and D), each bar represents the mean ± SD (n = 3). Mean data are normalized to β-Actin. (∗p < 0.05, ∗∗p < 0.01).

Fig 3: BCL6 is a negatively regulated transcription factor that regulates MFN2(A) Venn diagram of the four databases.(B) Predicted binding sites of BCL6 in the MFN2 promoter region.(C) Expression and quantification of BCL6 in HUVECs stimulated by Ang Ⅱ. Each bar represents the mean ± SD for triplicate experiments. Mean data are normalized to GAPDH (∗∗∗p < 0.001).(D) Dual fluorescence reporter gene assay was used to verify that BCL6 inhibited the transcriptional activity of MFN2. Each bar represents the mean ± SD (∗∗∗p < 0.001).(E) The protein expression and (F) quantification of MFN2, P53, and P21 in HUVECs after siBCL6. Each bar represents the mean ± SD for triplicate experiments. Mean data are normalized to GAPDH (∗∗p < 0.01, ∗∗∗p < 0.001).(G) The protein expression and (H) quantification of MFN2, P53, and P21 in HUVECs after ovBCL6. Each bar represents the mean ± SD for triplicate experiments. Mean data are normalized to GAPDH (∗∗p < 0.01, ∗∗∗p < 0.001).

Fig 4: BCL6 regulates endothelial cell senescence through MFN2(A) Protein expression levels and (C) quantification of P53, P21, and other genes after siMFN2 and siBCL6.(B) Protein expression levels and (D) quantification of P53, P21, and other genes after ovMFN2 and ovBCL6. In (C and D), each bar represents the mean ± SD for triplicate experiments (∗∗p < 0.01, ∗∗∗p < 0.001; #p < 0.05, ##p < 0.01).

Fig 5: Antitumor effects of WK369 on SKOV3 xenograft mouse models. (A) SKOV3 xenograft mice were administered WK369 (25 mg/kg/d) and WK369 (50 mg/kg/d), with FX1 (50 mg/kg/d) as a control (n = 7 mice per group). (B) Tumour volumes (mm3) evaluated once every 3 days for a total of 21 days. (C) Images of IHC staining of Ki67 are shown. Scale bars, 50 μm. (D) Body weights changes of each group on day 21. (E) mRNA expression of the BCL6 target genes from tumors was measured by RT-PCR assays. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, n.s. not significant by log-rank (Mantel-Cox) test.