Fig. 6
From: Integrin beta3 regulates clonality and fate of smooth muscle-derived atherosclerotic plaque cells
Integrin β3 in bone marrow-derived cells regulates SMC clonality. a, b Itgb3(−/−), ApoE(−/−), SMMHC-CreERT2, ROSA26R(Rb/+) mice were fed a HFD for 6 weeks. In a aortic root sections were stained with DAPI and directly imaged for Rb colors. Note, multiple patches of smooth muscle-derived plaque cells of different colors with limited mixing contrasts with monoclonal plaques in Itgb3(+/+) mice (Fig. 2a, b). In b percentage of labeled cells of each color in media and plaque is shown; n = 3 mice, 6 plaques. Chi-square test was used; NS, not significant. c Percent of plaque cells at 6 weeks of HFD labeled by any color were compared in ApoE(−/−), SMMHC-CreERT2, ROSA26R(Rb/+) mice that were also Itgb3 wild type or null. Itgb3(+/+) data is also shown in Fig. 2d. d–f ApoE(−/−), SMMHC-CreERT2, ROSA26R(Rb/+) mice were induced with tamoxifen, lethally irradiated, and transplanted with Itgb3(+/+), ApoE(−/−) (d) or Itgb3(−/−), ApoE(−/−) (e) bone marrow prior to HFD. In d, e after 12 weeks of HFD, aortic root sections were stained with DAPI and imaged for Rb colors. In f mice were fed a HFD for 6–12 weeks, and percentage of labeled cells of each Rb color in media and plaque were quantified; n = 5 recipient mice, 6 plaques for each donor genotype. g–j ApoE(−/−) SMC were cultured in medium conditioned by bone marrow-derived macrophages from Itgb3(+/+), ApoE(−/−) or Itgb3(−/−), ApoE(−/−) mice for 0 and 8 h or 48 h. In g, h bright-field images and percent of SMC coverage at 8 h of uncovered area at 0 h are shown, respectively; n = 4. In i, j EdU was added to culture medium for the last 8 h of the 48 h incubation, and then SMCs were stained for EdU and DAPI and percent of cells expressing EdU was quantified; n = 4. Student’s t-test was used. k Schematics of plaques in ApoE(−/−), SMMHC-CreERT2, ROSA26R(Rb/+) mice that are Itgb3 wild type or null were induced with tamoxifen, transplanted with ApoE(−/−) bone marrow fed a HFD for 12 weeks. Error bars represent standard deviations. Med, tunica media; Pl, plaque. Scale bars, 25 μm (a, d, e, i), 100 μm (g)
