Fig 1: Effect of Pb on protein levels of three V-ATPase subunits in isolated lysosomes of rPT cells. Cells were treated with Pb (0.25, 0.5 and 1 μM) for 12 h, then fractionated into the lysosomal lysis to assess the protein levels of ATP6V1A (a), ATP6V1B1+ATP6V1B2 (b) and ATP6V1D (c). Upper panels represent western blot images; lower panels represent quantitative analysis of protein levels (mean±S.E.M., n=4). ns, not significant, *P<0.05, **P<0.01, as compared with control
Fig 2: Low-dose celecoxib induced new bone formation in porous endplates. a Quantitative analysis of OSX mRNA expression in lumbar endplates at 2 and 4 weeks after celecoxib or vehicle treatment by qRT-PCR. b Representative images of immunostaining of osterix (green) and DAPI (blue) in the L4-L5 caudal endplates at 2 weeks after celecoxib or vehicle treatment. c Quantitative analysis of the percentage of osterix+ area in lumbar endplates. d Representative images of immunostaining of CD31 (green), endomucin (red), and DAPI (blue) in the L4-L5 caudal endplates at 2 weeks after celecoxib or vehicle treatment. e Quantitative analysis of the number of CD31hiendomucinhi cells in lumbar endplates. f Representative images of TRAP staining (magenta) in the L4-L5 caudal endplates at 2 weeks after celecoxib or vehicle treatment. g Quantitative analysis of the number of TRAP+ cells in lumbar endplates. h Quantitative analysis of Vpp3 mRNA expression in lumbar endplates at 2 and 4 weeks after celecoxib or vehicle treatment by qRT-PCR. i Representative images of immunostaining of Vpp3 (red) and DAPI (blue) in the L4-L5 caudal endplates at 2 weeks after celecoxib or vehicle treatment. j Quantitative analysis of the percentage of Vpp3+ area in lumbar endplates. Scale bars, 50 μm (b, d, f, i). *P < 0.05 compared with the sham group and #P < 0.05 compared with the vehicle group. n = 3 per group (a, h); n = 6 per group (c, e, g, j)
Fig 3: Recombinant Dll4(E12) increases osteogenesis.(A) Tile scan confocal images of Osterix (OSX) staining (green) in femurs from pLIVE-Dll4(E12) and control-injected mice. Nuclei, DAPI (blue). Graph shows quantitation of OSX+ cells. Data represent mean ± s.e.m. (n = 4 mice) (p-values determined by two-tailed unpaired t-test). (B) Runx2 staining (green) of pLIVE-Dll4(E12) and control femurs. Nuclei, DAPI (blue). Graph shows quantitation of Runx2+ cells. Data represent mean ± s.e.m. (n = 4 mice) (p-values determined by two-tailed unpaired t-test). (C) Representative images showing Osteopontin (Opn) and Osteocalcin staining (green) in pLIVE-Dll4(E12) and control femur. Nuclei, DAPI (blue). Data represent mean ± s.e.m. (n = 3 mice) (p-values determined by unpaired t-test with Welch’s correction). (D) Tile scan images of ATP6V1B1+ ATP6V1B2 (V-ATPase) staining (green) in pLIVE-Dll4(E12) and control femur. Nuclei, DAPI (blue). Graphs show quantification of osteoclast surface/bone surface (Os. S/B S) and osteoclast number/bone perimeter (No. Oc./B. Pm). Data represent mean ± s.e.m. (n = 4 mice) (p-values determined by unpaired t-test with Welch’s correction). (E) Tile scan images of Perilipin staining (green) in pLIVE-Dll4(E12) and control femurs. Nuclei, DAPI (blue). Graph shows quantitation of Perilipin+ adipocytes. Data represent mean ± s.e.m. (n = 3 mice) (p-values determined by unpaired t-test with Welch’s correction). Figure 3—source data 1.Source data for Figure 3A–E.
Supplier Page from Abcam for Anti-ATP6V1B1 + ATP6V1B2 antibody [EPR16401]