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. 2020 May 27;21(11):3774. doi: 10.3390/ijms21113774

Figure 4.

Figure 4

Absence of IL1-RA induces the expression of osteoclast markers particularly in long bone and jaw-derived osteoclasts. Gene expression was measured in wild-type (WT) and Il1rn−/− osteoclasts derived from long bone, calvaria, vertebra, and jaw bone marrow (BM) that were cultured on bone slices for 6 days. (A) Cultures of Il1rn−/− osteoclasts from all skeletal sites show no expression of Il1rn. The absence of Il1rn significantly increased the mRNA expression of Il1b in long bone and vertebra-derived osteoclasts (n = 6 mice/ group). (B) Protein levels of IL1-β were determined in the culture supernatants at day 3 of osteoclastogenesis. Whereas IL1-β production was below the detection limit in WT and Il1rn−/− osteoclasts from calvaria and jaw, Il1rn−/− osteoclasts derived from long bones and vertebrae showed increased IL1-β levels (n = 3 mice/group). nd = not detected. (C) Of note, the mRNA expression of various specific osteoclast markers was significantly increased exclusively in Il1rn−/− osteoclasts derived from long bone and jaw BM (n = 6 mice/group). Acid phosphatase 5 (Acp5); nuclear factor of activated T cells 1 (Nfatc1); dendritic cell-specific transmembrane protein (Dcstamp); cathepsin K (Ctsk). All values were normalized for the expression of Beta 2 microglobulin (B2m) as the reference gene. Relative expression is shown (* = p < 0.05, ** = p < 0.01, *** = p < 0.001 compared to WT controls).