Fig. 6
Perturbation to the intercellular network of a multicellular D3 ESC population affects RA-accelerated differentiation in a temporal manner. At 24 h, neither adenylyl cyclase (AC) inhibition (a, SQ-treatment) nor gap junction (GJ) inhibition (b, β-GA-treatment) induced a significant change in spatial or temporal characteristics of differentiation compared to the vehicle control. By 48 h, a temporal shift along PC1 is observed for both treatments (a, b), depicting a decrease in the rate of differentiation. The intercellular model accurately predicted these dynamics, represented by average values for SQ-treatment (Exp: n = 87, Sim: n = 30) and BGA-treatment (Exp: n = 62, Sim: n = 30) compared to the vehicle control (Exp: n = 87, Sim: n = 75) (a, b). A schematic diagram of our proposed mechanism for the influence of AC and GJ inhibition on differentiation potential is depicted in c. Specifically, both AC and GJ inhibition are suggested to decrease the intercellular flux between cells but via separate mechanisms: modulating the concentration gradient and the number of open channels for AC and GJ inhibition, respectively. Data clouds of a, b before averaging are shown in Supplementary Figures 8, 9
