Fig. 3

NR5A2 controls stemness in PDAC. A Sphere formation capacity on day 7 following Cpd3 treatment. Representative images for PDX215 and PDX354 following treatment with Cpd3 (60 µM). B Quantification of primary sphere formation capacity on day 7 of Cpd3 treatment for PDX215 (n = 4 biological replicates) and PDX354 (n = 10 biological replicates). Data are shown as violin plots with dotted lines indicating the median. C The schematic (created with biorender.com) illustrates the process of forming first-generation spheres over 7 days. This is succeeded by the disintegration of the formed spheres into a single-cell suspension, and then the commencement of second-generation spheres over another 7-day period, with or without Cpd3 treatment (upper panel). The lower panel shows the secondary sphere formation of cells treated with Cpd3 (60 µM) for five different PDAC cultures. D Colony formation capacity of cells treated with Cpd3 for 72 h. Representative pictures (top), quantification of colony formation (bottom). E Secondary sphere formation capacity following knockdown or overexpression (OE) of NR5A2 for PDX215 and PDX354. In panels C, D, and E data are presented as mean ± SD and statistically analyzed using two-tailed Mann–Whitney tests to compare two groups (n = 4 biological replicates). Asterisks indicate significance at the indicated levels: * p < 0.05, *** p < 0.001, and **** p < 0.0001. Please also see Supplementary Fig. 3