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. 2014 Oct 2;9(10):e109078. doi: 10.1371/journal.pone.0109078

Figure 3. Adseverin D5 is not is expressed during in vitro OCG.

Figure 3

A) PCR detected only the full-length Ads isoform in Day 0 and Day 6 BMMs and Day 0 and Day 4 RAW macrophage cultures. pSE380-Ads (full-length Ads) and pSE380-Ads-D5 (Ads D5 isoform) plasmids were used as positive controls. Equal volumes of cDNA from each representative samples were used as templates. The intensity of the signal is not a quantitative measure of Ads gene expression. B) Immunoblot analysis also failed to detect the Ads D5 in BMM and RAW macrophage-derived osteoclasts. Lysates from the transformed BL21 E. coli induced to express exogenous protein were used as positive controls. Only the 85 kDa full-length isoform was detected in 20 µg of TCL from Day 6 BMMs and Day 4 RAW macrophages cultures. Ads was not detected in 20 µg of TCL from unstimulated samples.