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. 2009 Apr 24;284(17):11531–11545. doi: 10.1074/jbc.M808324200

FIGURE 2.

FIGURE 2.

Cell type specificity of the pGL3-TRACP-1C-PTP-oc expression plasmid in vitro (A) and in vivo (B). A, an osteoclastic cell line (murine RAW264.7 monocytic cells) and four nonosteoclastic cell lines (murine MC3T3-E1 osteoblastic cells, human TE85 osteosarcoma cells, human HepG2 hepatic cells, and human HEK293 embryonic kidney cells) were transfected with pGL3-TRACP-1C-PTP-oc or pGL3-TRACP-1C empty plasmid (negative control). To determine relative transfection efficiencies in these cell lines, parallel cultures of each cell type were transfected with the pGL3-control plasmid. Relative rabbit PTP-oc mRNA levels and luciferase activity in each transfected cell cultures were determined as described under “Experimental Procedures.” To adjust for the vast differences in transfection efficiencies, the results are shown as relative -fold PTP-oc mRNA levels per luciferase activity (mean ± S.E. n = 4). None of the cell lines transfected with the pGL3-TRACP-1C empty plasmid showed detectable rabbit PTP-oc mRNA levels. B, total RNA was isolated from marrow-derived osteoclasts (OCL) and the spleen, kidney, liver, and heart of young adult male transgenic mice. Relative rabbit PTP-oc expression levels (normalized against respective β-actin mRNA levels) were determined by real time RT-PCR in triplicate as described under “Experimental Procedures.” Results are shown as the mean of relative -fold level of the normalized PTP-oc mRNA levels in the heart (i.e. the adjusted rabbit PTP-oc mRNA in hearts of transgenic mice was set as 1-fold).