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. 2023 May 4;42(12):e112712. doi: 10.15252/embj.2022112712

Figure 2. STING rapidly traffics to the Golgi and exits via vesicular trafficking.

Figure 2

  • A
    Sting −/− iBMDMs expressing mRuby3‐STING (magenta) and eGFP‐Golgin84 (green) were imaged live on the 3i marianas spinning disk microscope. Z stack images were acquired either before (i.e. untreated) or after 50 μg/ml DMXAA treatment (i.e. image captured at 12 min post DMXAA treatment). Data are shown as a maximum intensity projection (MIP) of Z stack images. Scale bar = 5 μm.
  • B
    Increase in Golgi accumulation of eGFP signal starting from 2 min after 50 μg/ml DMXAA stimulation of eGFP‐STING iBMDMs and presented as Golgi/ER intensity in arbitrary units (a.u.) over time. Line of best fit is shown in red. Data are shown as the mean ± SD (N = 8 cells) and representative of three individual experiments.
  • C
    eGFP intensity loss from the Golgi starting from 20 min after 50 μg/ml DMXAA stimulation of eGFP‐STING iBMDMs and presented as intensity of the Golgi (a.u.) over time. Line of best fit is shown in red. Data are shown as the mean ± SD (N = 12 cells) and representative of three individual experiments.
  • D
    Sting −/− iBMDMs expressing mRuby3‐STING were imaged live on the spinning disk microscope. Representative images of timelapse showing STING vesicles exiting the Golgi for a recording starting 27 min after 50 μg/ml DMXAA treatment. Data are shown as a MIP of Z stack images. Square region of interest (ROI) indicates zoomed insert. Scale bar = 5 μm. Corresponds to Movie EV4.
  • E
    eGFP‐positive small vesicle regions were counted for individual cells over time starting from 20 min after 50 μg/ml DMXAA stimulation of eGFP‐STING iBMDMs and presented as the number of endosomes per cell. Data are shown as the mean ± SD (N = 11 cells) and representative of three individual experiments.
  • F
    Sting −/− iBMDMs expressing eGFP‐STING were imaged live on the spinning disk microscope. A representative image of timelapse showing transport of STING vesicles for a recording starting 1 h 18 min after 50 μg/ml DMXAA treatment. Data are shown as a MIP of Z stack images. Square ROI indicates zoomed insert. Scale bar = 5 μm. Corresponds to Movie EV6.

Source data are available online for this figure.