Extended Data Fig. 9: STING co-localized with mNeonGreen-ubiquitin at Rab11-positive REs after stimulation.
a, mNeonGreen-ubiquitin (mNG-Ub, cyan) and mRuby3-STING (magenta) were stably expressed in Sting−/− MEFs. Cells were stimulated with DMXAA for the indicated times. Cells were then fixed, permeabilized, and immunostained with anti-GM130 (a Golgi protein, yellow) or anti-Rab11 (a recycling endosomal protein, yellow) antibodies. Scale bars, 10 μm, 1 µm in the magnified images. b, c, Fluorescence intensity profiles along the red lines in (a) are shown. d, The Pearson’s correlation coefficient between mNeonGreen-ubiquitin and GM130, or between mNeonGreen-ubiquitin and Rab11 are presented in box-and-whisker plots with the minimum, maximum, sample median, and first vs. third quartiles. e, Sting−/− MEFs reconstituted with mRuby3-STING were treated with indicated siRNAs. Cells were then incubated with DMXAA for 12 h. The fluorescence intensity of mRuby3-STING inside or outside Lamp1-positive compartments (Lamp1+) was quantified. Data are presented in box-and-whisker plots with the minimum, maximum, sample median, and first vs. third quartiles. f, Sting−/− MEFs stably expressing mRuby3-STING and Lamp1-EGFP were treated with indicated siRNAs. Cells were then incubated with DMXAA and E64d/pepstatin A for 12 h. The fluorescence intensity of mRuby3-STING inside or outside Lamp1-positive compartments (Lamp1+) was quantified. Data are presented in box-and-whisker plots with the minimum, maximum, sample median, and first vs. third quartiles. The sample size (n) represents the number of cells (d, e, and f). Source numerical data are available in source data.
