Supplemental Figure 1. Scatter plot of a PANC1 3D sphere CellTiterGlo assay plate. Each dot refers to the activity of a compound in a 1536-well microplate.

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Supplemental Table 1. Summary table of Curve Response Classes (CRC) for the MIPE qHTS

Supplemental Figure 1. Scatter plot of a PANC1 3D sphere CellTiterGlo assay plate. Each dot refers to the activity of a compound in a 1536-well microplate. A) % CV = 7 Z’= 0.69 X-axis refers to the 48 columns in a 1536-well plate, and y-axis refers to the relative luminescence units (RLU) from the CellTiterGlo assay. Column 1 is the no cell control; column 2,3 are the Borterzomib cell kill control; column 4 is the control with just DMSO added; and columns 5-48 corresponds to the sample field (see Materials and Methods for experimental protocol)

PANC1 Max Response <30% SN12C Max Response <30% Supplementary Figure 2. Target enrichment of pan active hits by cell line PANC1 Max Response <30% SN12C Max Response <30% -logp -logp Target Target PANC1 CRC -1.1 or -1.2 SNC12 CRC -1.1 or -1.2 -logp -logp Target Target

Supplemental Figure 3. Dose response curves for digoxin measuring % cell viability in the different cells and assay modes (●) PANC1 2D monolayers, (●) PANC1 3D Spheres, (●) PANC1 forming spheres, (○) SN12C 2D monolayers, (○) SN12C 3D spheres, and (○) SN12C forming spheres.

Supplemental Figure 4. Bright field images of PANC1 3D spheres PANC1 spheres, Flask, 5 days 400mm 1000mm PANC1 spheres, 384-well ULA round bottom plate, 5 days 400mm

Supplemental Figure 5. Fixed ratio dose responses of selected compound combinations on PANC1 3D Spheres generated in 384-well ULA round bottom plates using the CaspaseGlo reagent to measure apoptosis signal 16hrs 24hrs

Human Apoptosis Array: Supplemental Figure 6. Dot blots of Apoptosis protein antibody arrays. Protein arrays in each box were processed together as a set. See web link below for protein localization in the dot array. Digoxin+Navitoclax Navitoclax Digoxin Carfilzomib+Navitoclax Carfilzomib Carfilzomib+Bardoxolone Bardoxolone DMSO Human Apoptosis Array: https://resources.rndsystems.com/pdfs/datasheets/ary009.pdf

Supplemental Table 2. Fold-changes in apoptotic protein levels (measured as pixels in dot blot) in drug vs DMSO treated PANC1 3D Spheres, for 20 hrs.