Fig. 4: PKA inhibition in FD-SSCs suppresses osteoclastogenesis and promotes osteogenic differentiation and maturation.

a TRAP staining of tibias showing TRAP-positive cells in PKA, FD-PKI4A and FD-PKI mice. b ELISA results showing the serum TRACP-5b levels in PKA, FD-PKI4A and FD-PKI mice (Control n = 17, PKA n = 6, FD-PKI4A n = 5 and FD-PKI n = 13, biologically independent samples, one-way ANOVA with Tukey’s multiple comparisons test). c ELISA results of serum Rankl levels (Control n = 23, PKA n = 5, FD-PKI4A n = 5 and FD-PKI n = 14, biologically independent samples, one-way ANOVA with Tukey’s multiple comparisons test). d qPCR analyses of osteogenic markers in limb bones (Control n = 34, PKA n = 9, FD-PKI4A n = 7 and FD-PKI n = 31, biologically independent samples, two-way ANOVA with Sidak’s multiple comparisons test). e Immunohistochemical staining and f quantitative analyses showing increased Osx-positive cells in PKA and FD-PKI4A mice, which returned to baseline levels in FD-PKI mice (Control n = 11, PKA n = 9, FD-PKI4A n = 11 and FD-PKI n = 6, biologically independent samples, one-way ANOVA with Tukey’s multiple comparisons test). In contrast, Ocn-positive cells are decreased in PKA and FD-PKI4A mice but elevated in the FD-PKI mice (Control n = 15, PKA n = 15, FD-PKI4A n = 11 and FD-PKI n = 12, biologically independent samples, one-way ANOVA with Tukey’s multiple comparisons test). g EdU staining and h quantitative analysis demonstrating increased proliferation in PKA and FD-PKI4A mice, with no significant difference in proliferation between PKA mice and FD-PKI4A mice (Control n = 9, PKA n = 11, FD-PKI4A n = 8 and FD-PKI n = 13, biologically independent samples, one-way ANOVA with Tukey’s multiple comparisons test). The data are presented as box-and-whisker plots, with boxes representing the interquartile range (25th–75th percentiles), the minimum and maximum values reached by bars, the median plotted as a line in the middle, and the mean marked as “+”. Source data are provided as a Source Data file.