Fig 1: Spliceosome protein localization and expression in single mammalian cells. (a) The (sc)2WB assigns subcellular localization to cytoplasmic (TurboGFP, green; ß-tubulin, blue) and nuclear (SFPQ, magenta; PTBP1, gray) proteins, even when the targets are components of large molecular machines. The dashed line is the microwell border. A representative intensity profile and false-color micrograph are shown here. A dashed line denotes the microwell border in the intensity profile. (b) Bi-directional PAGE enhances selectivity because uni-directional PAGE cannot resolve ß-tubulin and SFPQ. The dashed line indicates the position of the next row of microwells (array period). Note that in the whole-cell scWB, the TurboGFP band has overrun into the next separation lane. (c) Mean-normalized expression (AUC/AUCmean) and (d) subcellular localization (Nuc=AUCnuclear/AUCtotal, Cyt=AUCcytoplasm/AUCtotal) of TurboGFP, ß-tubulin, SFPQ, and PTBP1 (n=44 cells). Error bars are±1 s.d. GFP, green fluorescent protein; PAGE, polyacrylamide gel electrophoresis; s.d., standard deviation.