Extended Data Fig. 1: Matrix stiffness mediates PDAC organoid clonal heterogeneity.
a, Schematic summarizing the protocol and potential outcomes of genetically barcoded PDAC organoid expansion within Cultrex and HELP matrices. b, Number of barcodes present within PDAC organoid populations expanded within Cultrex and HELP matrices for one to three passages (N = 3 independent biological replicates per matrix type, mean ± 95% confidence interval, center line represents linear fit for each matrix over three passages, shaded region represents 95% confidence interval of linear fit). Statistical analysis was performed using an ordinary one-way ANOVA with Tukey multiple comparisons correction between data points at passage three (****P < 0.0001 for comparison between HELP High and Cultrex/HELP Low, comparison between HELP Low and Cultrex was not significantly different). The initial Parent population was the same for all matrices (N = 1). c, Slope of linear fit from b for each matrix (N = 3 independent biological replicates per matrix type, mean ± 95% confidence interval, ordinary one-way ANOVA with Tukey multiple comparisons correction, ****P < 0.0001, NS, not significant). d, Frequencies of barcodes present in the indicated matrix at passage three. Barcodes are listed in rank order and colored bars correspond to barcodes unique to only the indicated matrix. The number of these unique barcodes and their cumulative frequency are reported for each matrix. e, Venn diagram of barcodes at passage three across indicated matrices. f, Correlation plots comparing barcode frequencies across Cultrex and HELP at passage three. Pearson r values are reported for each pairing. Dashed line represents perfect correlation. In d-f, only barcodes with a frequency >0.01% across all three biological replicates at passage three were included.
