Supporting Information.

Fig. A, Optimization of cell culture with conditioned medium for siRNA screening of J2 cells. I, Colony assays of HFKs after 5 days using conditioned medium (CM) from J2 cells in the absence or presence of Y-27632 (Y). II, Colony assays of HFKs grown in medium supplemented with 100%, 50%, 25% or no (FY) CM from J2 cells in the presence of Y. Cell growth was measured by sulforhodamine staining at 560 nm. Growth was proportional to the dilution of CM at 200 cells/well. Fig. B, Uncropped and unadjusted western blots shown in Fig. 1D. Fig. C, Analysis of growth and apoptosis in conditionally reprogrammed HFKs. HFKs were maintained in culture for 2 days with Y-27632 (Y), J2 cells (J2) or J2 cells and Y (J2+Y). I, FACS with propidium iodide (x-axis) and a FITC conjugated annexin V antibody (Y-axis). There was no significant change in the percentage of double-positive apoptotic cells (upper right quadrant). II, Western analysis shows the absence of cleavage of Caspase 9 and Caspase 3, which indicate a lack of active apoptosis. The absence of cleavage of LC3B (I) to LC3B (II) indicate a lack of autophagy. Fig. D, Network analysis of genes (highlighted in blue) whose knockdown decreased HFK growth. Fig. E, Uncropped and unadjusted western blots shown in Fig. 4C. Table A, Antibodies used for immunofluorescence and western blotting. Table B, The siRNA library targeting factors secreted by J2 cells. Table C, Gene profile associated with the effect of Y-27632 and J2 cells on HFKs. Table D, Reverse-Phase Protein Array (RPPA) analysis.
(PDF)