Fig. 4: Mfn2 binding to the AJ complex protein β-catenin is increased following inflammatory stimulation. | Nature Communications

Fig. 4: Mfn2 binding to the AJ complex protein β-catenin is increased following inflammatory stimulation.

From: Mitofusin-2 stabilizes adherens junctions and suppresses endothelial inflammation via modulation of β-catenin signaling

Fig. 4

Confluent HLMVECs were treated with TNFα (10 ng/mL) for 6 h. a Mfn2 was immunoprecipitated with Mfn2 specific antibody followed by Western blotting for β-catenin or Mfn2 antibodies. Uncropped blots can be found in the Source Data file. b The band intensities for Mfn2 and β-catenin interactions in (a) were quantified with ImageJ. Data are mean values ± SEM for three independent experiments. *p = 0.0403 by unpaired, one-tailed t-test. c HLMVECs expressing GFP or GFP-Mfn2 were stimulated with TNFα (10 ng/mL) for 6 h. The GFP-Mfn2 was immunoprecipitated with GFP-trap magnetic agarose followed by Western blotting for β-catenin and Mfn2 antibodies. Uncropped blots can be found in the Source Data file. d The band intensities of the Western blot were quantified with ImageJ. Data are mean values ± SEM for three independent experiments. *p = 0.0097 by unpaired, one-tailed t-test. e–h Proximity ligation assay (PLA). e The interaction of endogenous VE-cadherin and β-catenin in HLMVECs was examined using mouse VE-cadherin antibody and rabbit β-catenin antibody for positive control. The red dots show an interaction between VE-cadherin and β-catenin. f Quantification of the number of interactions (red dots) per field in (e). Data are mean values ± SEM from n = 6 for basal and n = 7 for TNFα treatment in independent biological replicates samples. **p = 0.0018 by paired, two-tailed t-test. g The interaction of endogenous Mfn2 and β-catenin in HLMVECs was examined with mouse Mfn2 antibody and rabbit β-catenin antibody. The green dots show interaction between Mfn2 and β-catenin, and DAPI (blue) indicates nuclei. h Quantification of the number of interactions (green dots) per cell in (g). Data are presented with box and whiskers plot and whiskers are Min to Max. n = 8 for basal and n = 14 for TNFα. ****p < 0.0001 by paired, two-tailed t-test. The images in (e) and (g) are representative from at least six independent biological replicates.

Back to article page