Fig 1: BMD is decreased in PD-L2-/-mice. (A) Femoral bone mineral density (BMD) evaluated by DXA of WT, PD-L2-/-, and PD-L1/L2-/- mice. BMD was significantly lower in PD-L2-/- mice compared with WT mice. (B) and (C) Representative µCT images through the distal femoral metaphysis from a WT and a PD-L2-/- mouse. (D) RANKL serum levels in WT, PD-L2-/-, and PD-L1-/-PD-L2-/- mice. Serum RANKL levels in WT mice were significantly lower than in knockout mice. (E) OPG serum levels in WT, PD-1-/-, PD-L2-/-, and PD-L1-/-PD-L2-/- mice. Serum OPG levels did not differ between WT and knockout mice. (F) The ratio between serum RANKL and OPG in WT, PD-1-/-, PD-L2-/-, and PD-L1-/-PD-L2-/- mice. The ratio did not differ between the mice. (n ?= ?4 in each group). Differences between the groups are evaluated by one-way ANOVA with subsequent t-tests. Bars represent mean with SD. Asterix (*) represents p ?< ?0.05 and ** represents p ?< ?0.01.
Fig 2: 5-MTP suppresses HFD-induced IL-6 and HSP60 in the atherosclerotic lesions of ApoE-/- mice. Chow diet-fed or HFD-fed ApoE-/- mice were treated with saline or L-5-MTP twice weekly for 20 wk. (A, B) IL-6 and (C, D) OPG in paraffin-embedded aortic tissues were analyzed by immunohistochemistry. Negative control represents staining with a 2nd antibody. Representative microphotographs of the immunohistochemical staining for (A) IL-6 and (C) OPG are shown. Immunopositive areas of (B) IL-6 and (D) OPG in aorta tissues were quantified as a percentage of the total aortic area in each section. Data represent mean ± SEM (Chow + saline group: n = 6, HFD + saline group: n = 6 and HFD + 5-MTP group: n = 5). E Plasma IL-6 levels in HFD-fed ApoE-/- mice with injection of saline (n = 19) or L-5-MTP (n = 9) twice weekly for 20 wk for were measured by ELISA. Data represent mean ± SEM. After pretreating VSMCs with DMSO, DL-5-MTP, or L-5-MTP (100 µM) for 30 min, VSMCs were treated with vehicle or Pam3CSK4 (Pam3) in calcifying medium for 24 h. F IL6 and G OPG level in culture supernatants was measured by ELISA. Data represent mean ± SD of at least 3 independent experiments. HSP60 contents in aorta tissues from ApoE-/- mice fed chow diet with saline (n = 5) or HFD with saline (n = 5) or L-5-MTP (n = 5) for 20 wk were analyzed by immunohistochemistry. Representative microphotographs of the immunohistochemical staining (H) and quantification of immunopositive positive areas (I) are shown. Data represent mean ± SEM. Lu lumen, Neo neo-intima, Med media, An adventitia. P < 0.05, P < 0.01 determined by a one-way ANOVA
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