Figure 8.
Localized optogenetic vimentin repositioning displaces dense ER matrices independent of RNF26. (A–C) U2OS WT (A) and RNF26 knockout (KO) (B) cells co-transfected with Vim-mCh-SspB, iLID-GFP-GCN4-ppKin14, and Halo-KDEL were pulsed with 488-nm light inside an ROI at the cell periphery (blue dashed box) for 20 min to locally displace vimentin. (A and B) Example images shown before blue light pulsing (0 min) and 5, 10, and 20 min after localized blue light application. The lack dashed box indicates the region of the cell enlarged in the zoom images. (C) Mean fluorescence intensity of Vim-mCh-SspB and Halo-KDEL (ER) inside the ROI pulsed with blue light was quantified over time and normalized to the first time point after subtracting background. Graph shows mean ± SD, n = 8–9 cells analyzed per group over three independent experiments.

Localized optogenetic vimentin repositioning displaces dense ER matrices independent of RNF26. (A–C) U2OS WT (A) and RNF26 knockout (KO) (B) cells co-transfected with Vim-mCh-SspB, iLID-GFP-GCN4-ppKin14, and Halo-KDEL were pulsed with 488-nm light inside an ROI at the cell periphery (blue dashed box) for 20 min to locally displace vimentin. (A and B) Example images shown before blue light pulsing (0 min) and 5, 10, and 20 min after localized blue light application. The lack dashed box indicates the region of the cell enlarged in the zoom images. (C) Mean fluorescence intensity of Vim-mCh-SspB and Halo-KDEL (ER) inside the ROI pulsed with blue light was quantified over time and normalized to the first time point after subtracting background. Graph shows mean ± SD, n = 8–9 cells analyzed per group over three independent experiments.

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