Fig. 3: PKA inhibitor peptide (PKI) suppresses PKA signaling.
From: Protein kinase A is a dependent factor and therapeutic target in mouse models of fibrous dysplasia
a Immunoblotting and b ELISA showing the phosphorylation levels of PKA and CREB in 293T cells stimulated with forskolin and 3-isobutyl-1-methylxanthine (FI) with or without PKI plasmid transfection (n = 6 independent experiments, two-way ANOVA with Tukey’s multiple comparisons test). Three independent immunoblotting were performed. c Schematic representation of the Tet-GαsR201C/Tet-PKI/Prrx1-Cre (FD-PKI mice) transgenic mouse model, which is engineered to express the GαsR201C mutation, thereby upregulating the Gαs/cAMP signaling pathway while concurrently expressing PKI to inhibit PKA signaling in a tissue-specific manner upon Dox administration. The Tet-GαsR201C/Tet-PKI4A/Prrx1-Cre (FD-PKI4A mice) model was generated via the same strategy. d qPCR analysis confirming PKI mRNA transcription in FD-PKI mice (Control n = 7 and FD-PKI n = 8, biologically independent samples, unpaired two-tailed t-test) and showing increased GNAS mRNA expression in limb bones of both FD-PKI and FD-PKI4A mice (Control n = 27, FD-PKI4A n = 7 and FD-PKI n = 12, biologically independent samples, one-way ANOVA with Tukey’s multiple comparisons test). e ELISA results showing cAMP levels in the serum of FD-PKI and FD-PKI4A mice (Control n = 6, FD-PKI4A n = 5 and FD-PKI n = 7, biologically independent samples, one-way ANOVA with Tukey’s multiple comparisons test). f Immunoblotting showing the phosphorylation of PKA in PKA, FD-PKI4A, and FD-PKI mice. 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.
