Figure 1.

Quality control pipeline for ER distribution under control conditions. (A) Random fields of view of ER-immunolabeled, untreated MCF-7 breast cancer cells automatically imaged at $20\times /0.75$, shown as maximum intensity projection after image deconvolution. Scale bar: $20\mathrm{\mu }\mathrm{m}$. (B) Heat map representation of the binned distribution of single cell ER levels across the indicated independent biological and technical replicates, with lower ER levels at the top and higher ER levels at the bottom of the heat map. (C) The distance between the ER distributions in two experiments was measured by earth mover’s distance (EMD), and the pairwise EMD distances were submitted to hierarchical clustering. The three main clusters “C1,” “C2,” and “C3” are indicated at the bottom of the heat map. (D) EMD distances calculated from the median ER distribution across all experiments in the main cluster from panel C. Experiments that deviated $>3\times$ standard deviations from the mean of the main cluster EMDs are identified according to the cluster map in panel C. The dotted line above $\text{EMD}=0.1$ represents three standard deviations from the mean of the main cluster (dotted line above approximately 0.05 EMD). Summary data for Figure 1B and D are found in Excel Tables S1 and S2, respectively. Note: EMD, earth mover’s distance; ER, estrogen $\text{receptor-}\mathrm{\alpha }$.