Skip to main content
. 2023 Mar 23;26(4):106470. doi: 10.1016/j.isci.2023.106470

Figure 2.

Figure 2

Dox. induction elicits osteoclast formation

(A) Representative images of ex vivo marrow cultures following Dox. induction (Gray = Phalloidin-Alexa488, Cyan = Hoechst).

(B) Quantification of the number of fusion events over time following addition of Dox. Each time course was normalized to the number of fusion events observed on day 5. (n = 3).

(C) qPCR evaluation of the osteoclastogenesis transcription factors Nfatc1 and cFos under uninduced (black), Dox. induced (gray) or Dox. induced +100 ng/mL OPG (white) conditions. Fold expression is relative to Actb RNA. (n = 3).

(D) Quantification of the number of fusion events in ex vivo marrow cultures treated as in (C). (n = 3).

(E) Representative Western blot evaluation of the steady-state level of the osteoclast resorption enzyme CTSK in ex vivo marrow cultures treated as in (C).

(F) Representative immunofluorescence images of the pan macrophage lineage marker CD68 (green), the osteoclast sorting nexin SNX10 and Hoechst in ex vivo marrow cultures treated as in (C).

(G) Quantification of bone resorption by ex vivo marrow cultures treated as in (C). In (A and D), Arrowheads = multinucleated osteoclasts, Arrows = proliferative preosteoblasts. Error bars = ±SEM. Statistical significance was assessed via paired t-test, where ∗p=<0.05, ∗∗p=<0.01, ∗∗∗p=<0.001.