Figure 5
From: Npp1 prevents external tooth root resorption by regulation of cervical cementum integrity
Extracellular ATP induces the expression of Rankl in cementoblasts. (a) Extracellular ATP was measured with conditioned media (CM) from the culture of OCCM-30 cells with shEnpp1 and shNC (n = 5). (b) Transcript levels of Rankl and Opg were analyzed by real-time qPCR (n = 3). RNA was isolated from OCCM-30 cells treated with ATP for 24 h. (c) Protein amount of soluble Rankl was measured by ELISA using conditioned media (CM) from the culture of OCCM-30 cells treated with ATP (n = 4). (d) Protein levels were analyzed by Western blotting using specific antibodies with whole cell lysates from OCCM-30 cells treated with ATP for 24 h. Samples shown from Western blotting are from the same experiment, and the gels/blots were processed under the same experimental conditions. β-Actin was used as a loading control. Cropped images are displayed here; the original full-size blots are presented in Supplementary Fig. S6 online. (e) The protein amount of Rankl was measured by ELISA using whole cell lysates from OCCM-30 cells pretreated with H89 for 1 h and then treated with ATP (50 μM) for 48 h (n = 4). Significance was assigned with p-values as indicated in each graph.
