Figure 4
From: Npp1 prevents external tooth root resorption by regulation of cervical cementum integrity
Inactivation of Enpp1 induces Rankl expression in cementoblasts. (a) NTPPPHase activity was analyzed with OCCM-30 cells after treatment with Enpp1 inhibitor C for 2 h as indicated (n = 6). (b) Transcript levels of Rankl and Opg were analyzed by real-time qPCR (n = 3). RNA was isolated from OCCM-30 cells treated with Enpp1 inhibitor C for 24 h. (c) The protein amount of soluble Rankl was measured by ELISA using conditioned media (CM) from the culture of OCCM-30 cells with shEnpp1 and shNC (n = 3). (d) The protein amount of Rankl was measured by ELISA using whole cell lysates from OCCM-30 cells with shEnpp1 and shNC (n = 4). (e) TRAP staining was performed with BMMs from WT and Enpp1asj mice. BMMs were differentiated for 9 days with conditioned media (CM) from the culture of OCCM-30 cells supplemented with M-CSF (20 ng/ml) but without recombinant Rankl for 9 days. Scale bar are indicated. (f) TRAP-positive multinucleated cells were randomly counted in the field of the microscope of (e) at day 9 (n = 6). (g, h) Transcript levels of Trap and CtsK were analyzed by real-time qPCR (n = 3). RNA was isolated from BMMs differentiated in the same condition with (f). Significance was assigned with p values as indicated in each graph.
