Figure. Single-cell GFP RNA and protein quantification. (a) GFP induction quantified by labeled mRNA fluorescence (upper
), RT-qPCR normalized to Oct4 (middle
), and protein fluorescence (lower
). (b) GFP induced (1 µg/mL doxycycline) and uninduced mESC mixture with indicated positive (red) and negative (black) sorting gates. (c) RT-qPCR (normalized to Oct4) for GFP mRNA in each sorted fraction. (d) Single cell correlation of GFP mRNA and protein fluorescence in iPSCs heterozygous for Oct4-IRES-GFP (iPS-2i; see the article's Supplementary Information at http://dx.doi.org/10.1038/nmeth.2910
). (e) Correlation and linear scaling of GFP protein with mRNA in single cells for representative doxycycline induction levels. GFP, green fluorescent protein; RT-qPCR, reverse-transcription quantitative polymerase chain reaction; mESCs, mouse embryonic stem cells; iPSCs, induced pluripotent stem cells.