Supplementary MaterialsSupplFigure 1: Body S1. 2014). Data are represented as Mean SD. NIHMS1607090-supplement-SupplFigure_1.pdf (976K) GUID:?30F10BFB-3F64-4B91-BE74-0F04E22A4F1D SupplFigure 3: Physique S3. Analytical screening identifies all stable two-cell circuits. (related to Physique 3) A-C. Schematic depicting possible interactions between two different cell types. The first situation is unstable because neither cell type is usually close to transporting capacity and therefore no amount of regulation will result in a stable ON state (A). The second situation, when SAR-7334 HCl both cell types are close to transporting capacity, is also stable and no additional regulation is required because the transporting capacities enforce an ON state (B). The last situation, where one cell type is usually close to transporting capacity and the other is not, requires that this growth factor of SAR-7334 HCl the cell far from transporting capacity undergoes unfavorable regulation in order to achieve a stable ON state (C).D. Hierarchical representation of the 24 stable circuits that possess all three stable says, OFF, ON, and ON-OFF. Circuits are grouped based on the total quantity of interactions. NIHMS1607090-supplement-SupplFigure_3.pdf (592K) GUID:?42784361-8AB7-4922-A2CA-836BDC33B0D2 SupplFigure 4: Physique S4. Carrying capacity is regulated by space, nutrients, and growth factor availability (related to Physique 4) A. EdU labeling of FB plated in 6-well or 12-well tissue culture plates at different cell densities, 320,000; 240,000; 160,000; 80,000; 40,000; 20,000 cells/well (6-well) and 160,000; 120,000; 80,000; 40,000; 20,000; 10,000 cells/well (12-well). FB were labeled with EdU for 2 hr after overnight culture in 2% FBS. Dashed collection indicates linear regression of the data (data representative of two impartial tests, n=2).B. EdU labeling of FB plated in 6-well tissues lifestyle plates at different cell densities, 320,000; 240,000; 160,000; 80,000; 40,000; 20,000 cells/well. FB had been tagged with EdU for 2 hr after right away lifestyle in either 2% or 10% FBS. Dashed series signifies linear regression of the info (data representative of two indie tests, n=2). C. MP:FB ratios 7 and 2 weeks after co-culture in either 2% or 10% FBS. Beginning cell numbers had been 10:1 (160,000 & 16,000), 1:1 (80,000 & 80,000), and 1:10 (16,000 & 160,000). (data representative of two indie tests, n=2). D. EdU labeling of MP activated with Csf1 at different cell densities. MP had been tagged with EdU for 2 hr following the addition of PBS or 50 ng/ml Csf1 for 16 hr. Dashed series signifies linear regression of the info. Histograms show types of EdU+ stream cytometry gating for every cell thickness (data representative of three indie tests, n=2). E. EdU labeling of FB activated with recombinant Pdgfb at different cell densities. FB had been tagged with EdU for 2 hr following the Bmp6 addition of PBS or 50 ng/ml Pdgfb for 16 hr. SAR-7334 HCl Dashed series signifies linear regression of the info. Histograms show types of EdU+ stream cytometry gating for every cell thickness (data representative of three indie tests, n=2). F. Ramifications of development elements on MP and FB. Carrying capability and proliferation price () of FB by adding 50 ng/ml Pdgfb, and proliferation price of MP with addition of 50 ng/ml Csf1 for 16 hr had been SAR-7334 HCl assessed after 2 hr of EdU labeling (pooled data from three indie tests). *p 0.05; ** p 0.005, students t-test. G. Surface area expression of development factor receptor assessed by stream cytometry. Representative histograms of Csf1r, Pdgfra, and Pdgfrb amounts on either MP or FB at 60 min post development factor arousal (solid lines) in comparison to unstimulated cells (dashed lines). H. The uptake of recombinant Csf1 by MP and recombinant Pdgfb by FB. 500,000 MP and 50,000 FB had been plated in 12-well plates. Prior to the assay, MP had been deprived of L929 moderate dietary supplement and FB had been deprived of FBS for 16.