Fig. 4
Overview of 3D acini formation screen. a Optimization of functional screen conditions. 3D growth of MCF-10A_Src cells transfected with siPLK1 (positive control for screen), siSrc (against the expressed Src Y527F) and controls (mock and NTP). Scale bar represents 600 μm. The size of >100 acini were measured and plotted on a box and whisker graph, with center line indicating the median, the upper and lower bounds of box indicating the 25th and 75th percentiles of the data, and the upper and lower whiskers indicating the 5th and 95th percentiles of the data. b Primary screen results for MAP4K5 (growth repressor) and MAP3K6 (required for acinar growth). 3D growth of MCF-10A_Src cells transfected with siSMARTpools as indicated. Scale bar represents 400 μm. The quantification method and representation was as for (a). c Validation of the primary hit SGK1 by SMARTpool deconvolution. 3D growth of MCF-10A_Src cells transfected with individual siRNAs from the SMARTpool (siRNA1-4) (left). Scale bar represents 600 μm. The quantification method and representation (middle) was as for (a). Lysates from cells in 2D culture transfected with the same siRNAs were Western blotted as indicated (right). a–c For box and whisker graphs *p < 0.05, **p < 0.01, ****p < 0.0001 by Mann–Whitney test
