Fig. 6

In-silico vs. In-vitro dose-response effect of PKA activation with FGFR1 inhibition. The most potent condition from the screening is investigated further for a potential dose effect. A Fold change (FC) in ALP activity, with respect to control, due to PKA activator (Forskolin, 1μM) or FGFR1 inhibitor (PD161560, 125nM, and 625 nM) or the combination of both. B A range of values for PKA and FGFR1 imposed activities is screened in silico with 0 meaning no activity and 1 being the max possible activity. The percentage of transitions remaining in the hypertrophic state or transitioning towards the healthy state is reported in the upper panels, the rest of the transitions go to the “None” state. In the middle panels, fold change (resp. inverse of fold change) in DNA-normalized ALP activity with respect to control DMSO in ATDC5 is reported for a range of Forskolin and PD161570 concentrations. The in-vitro situation without drugs (yellow rectangle) would correspond to the basal level of PKA and FGFR1 in in-silico hypertrophy but there is no one-to-one correspondence between the in silico and in vitro ranges. All in vitro results represent n=9 (3 bio-replicates in 3 independent experiments), p-values are computed on log-transformed data with a linear mixed-effect model, user-defined contrasts (only combination versus corresponding single doses were compared), one-sided test and adjustment for multiple comparisons with the Holm’s method. The combinatorial treatment effects were greater than the ones for either of the single treatment both in silico and in vitro, for all concentrations in the gradient of dose relationships investigated