Fig. 3: PRL2 deficiency enhances the canonical RANK-signaling pathway via Rac1 and ROS.
A Representative immunoblot images for GTP-Rac1 in BMMs isolated from Ptp4a2fl/fl mice (referred to as WT) and Ptp4a2fl/flLysMCre+ mice (referred to as KO) stimulated with RANKL for the indicated times. B Dynamic curves of ROS production (left panel) and area under curve (AUC, right panel) in WT and KO BMMs stimulated with RANKL (RLU, relative light unit). C Mean fluorescence intensity (MFI) of DCF-DA in WT and KO BMMs with or without RANKL for 30 min (left panel). Quantification of relative DCF level, normalized by WT BMMs without RANKL (right panel). D Representative immunoblot images for phosphorylated forms of signaling proteins in WT and KO BMMs stimulated with RANKL for the indicated times. E Dynamic curves of ROS production (left panel) and AUC (right panel) in Ptp4a2 knockout RAW 264.7 cells (referred to as KO) replenished with control (Ctrl) or PRL2 with RANKL. F Representative immunoblot images for NFATc1 in KO RAW 264.7 cells replenished with Ctrl or PRL2 with or without RANKL for 2 days. G Dynamic curves of ROS production (left panel) and AUC (right panel) in WT and KO RAW 264.7 cells replenished with Ctrl and Rac1-T17N with RANKL. H Representative immunoblot images of NFATc1 in WT and KO RAW 264.7 cells replenished with Ctrl and Rac1-T17N with or without RANKL for 2 days. The data are representative of two or three independent experiments (biological replicates), and for B, C, E, and G, the data are shown as the mean ± s.e.m., *P < 0.05, **P < 0.01, ***P < 0.005, unpaired two-tailed Student’s t-test.
