Figure 3.
PRDX1 induces expressions of Saa3 and Acod1 via TLR4 to inhibit osteoclast differentiation
(A) Volcano plot of transcriptomic changes in PRDX1 (PRDX1 + no RANKL condition) compared to control (no PRDX1 + no RANKL). Colored dots correspond to genes with significant (FDR <0.05) and greater than 1.5-fold expression changes. Baselines indicate genes with substantial (-log FDR >30) and greater than 2.5-fold expression changes.
(B) OCP cells were treated with M-CSF (30 ng/mL) in the presence or absence of PRDX1 for 1 day. Saa3 and Acod1 mRNA expressions were measured by RT-qPCR.
(C) OCP cells were incubated for 4 days with SAA3 at various concentrations in the presence of M-CSF (30 ng/mL) and RANKL (100 ng/mL). TRAP staining was performed for the counting of multinucleated cells. Scale bar, 75μm.
(D) OCP cells were incubated for 2 days with PRDX1 (200 nM) in the presence of RANKL (100 ng/mL). Cellular ATP levels were measured.
(E) Saa3 and Acod1 expression with TAK242 (5 μM) in the presence of PRDX1 (200 nM) for 1 day was measured by RT-qPCR.
(F) Saa3 and Acod1 expression with MyD or TRIF inhibitor (each 5 μM) in the presence of PRDX1 (200 nM) for 1 day was measured by RT-qPCR. The results were reported as Mean ± SD from three independent experiments. p value is determined by a two-tailed t-test in (D) and one-way ANOVA with Tukey’s multiple comparisons test in (B, C, E, and, F). ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001.