Fig. 4: Live-cell SIM imaging reveals the dynamics of organelles.

a SIM images of live U-2 OS cells after a 1-h coincubation with PV-1 (4 μM) and Tubulin-Atto 488, Tubulin-Atto 565, or Tubulin-Alexa 647 (5 μM). Additional frames are shown in movie S1. b Enlarged time-lapse images of the boxed region in (a). Representative images of consecutive SIM frames are displayed, and additional frames are shown in movie S2. c The cross-sectional profile of microtubules in (b) with full-width at half-maximum (FWHM) of 108 nm. d An SIM image of live U-2 OS cells expressing SNAP-Sec61β after a 1-h coincubation with PV-1 (4 μM) and SNAP-Alexa 488 (5 μM), and enlarged time-lapse images of the boxed region. Representative images of consecutive SIM frames are displayed, and additional frames are shown in movies S3 and S4. e A dual-color SIM image of live U-2 OS cells after a 1-h coincubation with PV-1 (4 μM), Tubulin-Atto 488 (5 μM), and Morph-Alexa 647 (3 μM). Additional frames are shown in movie S5. f, g Enlarged time-lapse images of the boxed regions in (e). Representative images of consecutive SIM frames are displayed, and additional frames are shown in movies S6 and S7. h A dual-color SIM image of live U-2 OS cells expressing SNAP-Sec61β after a 1-h coincubation with PV-1 (4 μM), SNAP-Alexa 488 (5 μM), and Morph-Alexa 647 (3 μM). Additional frames are shown in movie S8. i Enlarged time-lapse images of the boxed region in (h) reveal dynamic interactions between lysosomes and the ER. Representative images of consecutive SIM frames are displayed, and additional frames are shown in movie S9. j, k Representative time-lapse SIM images reveal different types of dynamic interactions between lysosomes and the ER in live U-2 OS cells expressing SNAP-Sec61β after a 1-h coincubation with PV-1 (4 μM), SNAP-Alexa 488 (5 μM), and Morph-Alexa 647 (3 μM). Additional frames are shown in movies S10 and S11. Scale bars: a, e, and h, 2 μm; b, d, f, g, i, j and k, 1 μm