Leishmania donovani restricts mitochondrial dynamics to enhance miRNP stability and target RNA repression in host macrophages

Supplemental Materials

This article contains the following supporting material:

  • Supplemental Materials
  • Video S1 - Supplementary Video 1. Mitochondrial movements in non-infected RAW264.7 macrophage cells. Time lapse live cell imaging of mitochondria in non-infected RAW264.7 cells imaged for 1 frame per second for 60 frames. Cells were transfected with a mitochondrial targeting variant of GFP (MT-GFP, green). The video plays at 15 frames per second. Scale bars, 5 μm. Time is in hours:minutes:seconds. Also see Supplementary Figure 2.
  • Video S2 - Supplementary Video 2. Mitochondrion movements in Ld infected RAW 264.7 macrophage cells. Time lapse live cell imaging of mitochondria in Ld infected RAW 264.7 cells imaged for 1 frame per second for 60 frames. Cells were transfected with a mitochondrial targeting variant of GFP (MT-GFP, green). Ld were stained using Mitotracker red (red) for 30 minutes and Raw macrophages were infected with Mitotracker red pre-stained Ld for 24 hrs before the time lapse imaging. The video plays at 15 frames per second. Scale bars, 5 μm. Time is in hours:minutes:seconds. Also see Supplementary Figure 2.
  • Video S3 - Supplementary Video 3. Time-lapse imaging of ER-endosome dynamics in Wild type MEF cells. Time lapse imaging of ER and EE in MEFs cells imaged for 1 frame per second for 115 frames. Cells have been co-transfected with pDsRed2-ER (red) and pAcGFP1-Endo (green) plasmids. The video plays at 15 frames per second. Scale bars, 10 μm. Time is in hours:minutes:seconds. Also see Supplementary Figure 6.
  • Video S4 - Supplementary Video 4. Time-lapse imaging of ER-endosome dynamics in Mfn2 -/- MEF cells. Time lapse imaging of ER and EE in Mfn2 -/- MEFs cells imaged for for 1 frame per second for 115 frames. Cells have been co-transfected with pDsRed2-ER (red) and pAcGFP1-Endo (green) plasmids. The video plays at 15 frames per second. Scale bars, 10 μm. Time is in hours:minutes:seconds. Also see Supplementary Figure 6.
  • Video S5 - Supplementary Video 5. Mitochondrial FRAP dynamics in non-infected RAW264.7 macrophage cells. Time lapse live cell imaging of mitochondria showing FRAP recovery of Mito-GFP in non-infected Raw 264.7 cells imaged at 2 frame per second for 220 frames post photobleaching. Cells were transfected with a mitochondrial targeting variant of GFP (Mito-GFP, green). The video plays at 15 frames per second. Scale bars, 5 μm. Time is in hours:minutes:seconds.
  • Video S6 - Supplementary Video 6. Mitochondrial FRAP dynamics in Ld infected RAW 264.7 macrophage cells. Time lapse live cell imaging of mitochondria showing FRAP recovery of Mito-GFP in 24h Ld infected Raw 264.7 cells imaged at 2 frame per second for 220 frames post photobleaching. Cells were transfected with a mitochondrial targeting variant of GFP (Mito-GFP, green). The video plays at 15 frames per second. Scale bars, 5 μm. Time is in hours:minutes:seconds.
  • Video S7 - Supplementary Video 7. Mitochondrial FRAP dynamics in Ld infected RAW 264.7 macrophage cells treated with Genipin (100μM). Time lapse live cell imaging of mitochondria showing FRAP recovery of Mito-GFP in 24h Ld infected Raw 264.7 cells treated with 100μM Genipin 4h post Ld infection, imaged at 2 frame per second for 220 frames post photobleaching. Cells were transfected with a mitochondrial targeting variant of GFP (Mito-GFP, green). The video plays at 15 frames per second. Scale bars, 5 μm. Time is in hours:minutes:seconds.