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. Author manuscript; available in PMC: 2015 Oct 23.
Published in final edited form as: Dev Biol. 2008 Oct 25;325(1):129–137. doi: 10.1016/j.ydbio.2008.10.014

Fig. 3.

Fig. 3

Expression of WT- and MT-DLX3. (A) Western blot analyses performed using the 6095 antibody that detects both WT- and MT-DLX3 proteins, C-20 antibody that detects WT-DLX3 protein, and a MT-DLX3 specific antibody in MC3T3E1 cells transfected with empty vector (EV), WT-DLX3, and MT-DLX3. (B) Real time PCR analysis demonstrating DLX3 mRNA expression in primary osteoblasts from Transgenic (TG) compared to control (Con) littermates. Fold increase represents the ratio of 2^-(Ct(Target)-Ct(GAPDH))(TG)/2^-(Ct (Target)-Ct(GAPDH))(Con) (p: * <0.05, data represent mean±SEM). (C) Primary osteoblasts from TG mice expressing both WT- and MT-DLX3 proteins and cells from control littermates expressing only WT-DLX3 protein (left panel). Quantification of WT- and MT-DLX3 proteins in primary osteoblasts (open bar; WT-DLX3, hatched bar; MT-DLX3) (right panel). (D) Immunohistochemical detection of WT- and MT-DLX3 proteins in bone sections from 6 week old male MT-DLX3 TG mice. WT-DLX3 protein but not MT-DLX3 protein was detected in of osteoblasts as indicated by arrowheads and osteocytes as indicated by the arrows in control littermates. TG mice expressed both WT- and MT-DLX3 proteins in the nucleus of osteoblasts (arrow heads) and osteocytes (arrows). Higher magnification shows localization of WT- and MT-DLX3 proteins in nucleus (inserted images). Pre-bleed serum was used as a negative control (Pre Ab). (E) Immunofluorescence double labeling demonstrating the WT-DLX3 (red) (c) and MT-DLX3 (b) proteins expressed in the nucleus of primary osteoblasts from TG mice (d; merge, a; phase contrast).