Fig. 1: PKAcα^W197R mutation enhances PKA signaling.

a Luciferase assay showing transcriptional activation of cAMP-responsive element (CRE) was elevated by both PKAcα and its constitutively active mutant PKAcα^W197R in 293T cells (n = 9 independent experiments, one-way ANOVA with Tukey’s multiple comparisons test). PKAcα^W197R induced more robust activation of CRE luciferase activity than wild-type PKAcα. b Immunoblot showing the phosphorylation of PKA and CREB in PKAcα^W197R-transfected cells, with HA-tag detection confirming transfection. Three independent experiments were performed. c ELISA results showing the level of phosphorylated CREB in PKAcα^W197R-transfected cells (n = 6 independent experiments, two-way ANOVA with Sidak’s multiple comparisons test). d Schematic of the Tet-PKAcα^W197R/Prrx1-Cre transgenic mouse model (PKA mice) for tissue-specific expression of PKAcα^W197R upon Dox administration. e qPCR analysis of PRKACA expression in limb bones of PKA mice (Control n = 5 and PKA n = 8, biologically independent samples, unpaired two-tailed t-test). f Immunoblotting confirming phosphorylated PKA levels in limb bones. Three independent experiments were performed. 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.