Fig 1: a Detection of PTHrP expression in DFC_B by immunofluorescence. Overlay of transmitted light and fluorescence image. The red bar corresponds to 50 µm. Images were obtained with a 40 × magnification objective lens. Exposure time was 0.02 s for transmitted light image and 2 s for fluorescence image. b Gene expression (real-time RT-PCRs) of PTHrP (PTHLH), BMP2 and ALP (ALPL) in DFC_B after gene silencing of PTHrP. For control, an unspecific siRNA (AllStars) was used. Columns represent mean + SD (n = 3), *p < 0.05. c Relative degree of the phosphorylation of the SMAD1/5 protein (western blot analyses) in DFC_B after gene silencing of PTHrP. For control, an unspecific siRNA (AllStars) was used. Columns represent mean + SD (n = 3). Whole length blots and a total lane protein image can be found in Supplementary Figure S1
Fig 2: a ALP activity in DFC_B after 7 days treatment with PTHrP or with the PTH receptor inhibitor (Inh) PTH (7–34) and osteogenic differentiation. Columns represent mean + SD (n = 3). b ALP activity in DFC_B after gene silencing of PTHrP with specific siRNAs after 7 days of osteogenic differentiation. For control, an unspecific siRNA (AllStars) was used. Columns represent mean + SD (n = 3), *p < 0.05. c Estimation of cell proliferation of DFC_B with CCK8 assay after inhibition of PTHrP gene expression. Columns represent mean + SD (n = 3)
Fig 3: a Relative gene expression (real-time RT-PCRs) of osteogenic differentiation markers in DFC_A and in DFC_B. Columns represent mean + SD (n = 3), *p < 0.05. b Relative degree of the phosphorylation of the SMAD1/5 protein (western blot analyses) and relative gene expression of BMP2 (real-time RT-PCRs) in DFC_A and in DFC_B. Columns represent mean + SD (n = 3), *p < 0.05. Whole length blots and a total lane protein image can be found in Supplementary Figure S1. c Relative amounts of extracellular PTHrP in cell culture media of DFC_A and DFC_B. DFCs were cultivated in standard medium (DMEM) or in osteogenic differentiation medium (ODM). Columns represent mean + SD (n = 3), *p < 0.05
Supplier Page from Abcam for Anti-PTHLH antibody