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. Author manuscript; available in PMC: 2018 Feb 27.
Published in final edited form as: Sci Signal. 2017 Dec 12;10(509):eaan6282. doi: 10.1126/scisignal.aan6282

Figure 4. Knockdown of CD32 in myeloma cells reduces CRP-induced OC differentiation.

Figure 4

(A) Western blot analysis for the abundance of CD32 in cultured wild-type (wt) ARP-1 or MM.1S cells and those transfected with non-targeted shRNA (Ctrl) or CD32 shRNA (CD32-KD). β-actin served as loading control. (B) The number of multinuclear TRAP+ cells in cocultures of OC precursors with control or CD32-deficient myeloma cells in the presence of CRP (5 μg/mL) or PBS. (C) Real-time PCR of gene expression in OCs cocultured with control or CD32-deficient ARP-1 myeloma cells and CRP (5 μg/mL) or PBS. (D to G) Histomorphometric analysis of (D) bone volume (BV/TV), (E) the number of OCs present on the bone surface (Oc. S/BS), and immunohistochemcial examination for (F) the percentage of CD138+ cells (myeloma marker) in the bone marrow, and (G) staining for CD32 and CD138 by myeloma cells in bone sections of distal femurs from SCID mice (n=10 per group) injected with Ctrl or CD32-KD ARP-1 cells. Mice receiving no CRP injection served as control in (D) and (E). Scale bar, 50 μM. *P < 0.05, ** P < 0.01 by student’s t test. Data from three independent experiments are shown.