Fig. 5

Inhibition of the PD-1/PD-L1 signaling during early osteoclast differentiation leads to a decrease in osteoclastogenesis. a Representative TRAP⁺ images after PD-1 and PD-L1 inhibition during days 0-3 of osteoclastogenesis (scale bar = 0.5 mm). b Representative TRAP⁺ images after PD-1 and PD-L1 inhibition during days 3-6 of osteoclastogenesis (scale bar = 0.5 mm). c Number of TRAP⁺ cells after PD-1 and PD-L1 inhibition during days 0-3 and days 3-6 (n = 8; each dot represents an averaged technical replicate). d Average diameter (μm) of TRAP⁺ cells after PD-1 and PD-L1 inhibition during days 0-3 and days 3-6. e Gene expression of Ostm1, an osteoclast lineage commitment gene, relative to β-Actin after PD-1 and PD-L1 inhibition during days 0-3 and days 3-6 (n = 6). f Gene expression of c-FOS, an osteoclast lineage commitment gene. g Gene expression of Dc-stamp, an osteoclast fusion gene. h Gene expression of Dc-stamp, an osteoclast fusion gene. i Gene expression of Ctsk, an osteoclast resorption gene. j Gene expression of Acp5, an osteoclast resorption gene. Analyses for c, d were performed as a 2-way ANOVA with Šídák’s multiple comparisons test. Analyses for e–i were performed as a 2-way ANOVA with Tukey’s multiple comparisons test