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. 2021 Mar 5;12:1463. doi: 10.1038/s41467-021-21626-1

Fig. 5. Kinesin-1 activity is required for QPD crystal formation in living cells.

Fig. 5

a U2OS transfected with Kin330-GFP plasmid and treated with QPD-OTf (20 µM, 2.5 h); green: crystals, magenta: Kin330-GFP. b U2OS transfected with Kin560-GFP plasmid and treated with QPD-OTf (20 µM, 2.5 h) (left); zoom of highlighted boxes, arrows indicate stabilized MTs correlating with crystals (right); green: crystals, magenta: kinesin. Scale bar: 50 µm. c Quantification of crystal formation in transfected cells vs control; n = 20; an average of three independent experiments; data are presented as mean value ± the standard error of the mean (SEM). Statistics were calculated using a two-tailed t-test; **p = 0.0064 (Control vs Kin330), **p = 0.0024 (Kin330 vs Kin560); nsp = 0.065. d Kinesin-1 knockdown experiment. Representative images of HeLa-GFP-Tubulin treated with RNA control sequence + QPD-OTf (20 µM, 2 h) (top) or with kinesin-1 siRNA + QPD-OTf (20 µM, 2 h) (bottom). Scale bar 100 µM. e Quantification of crystal intensity for the kinesin-1 knockdown experiment. a.u. represent arbitrary units; n = 30; data are the average of three independent experiments; data are presented as mean value ± the standard deviation (SD); statistics were calculated using a two-tailed t-test; ***p = 0.0007. f Kif5B and Tubulin bands from western blot assay for the kinesin-1 knockdown experiment in HeLa cells. Samples derive from the same experiment and blots were processed in parallel. g U2OS treated with kinesore (100 µM) in Ringer’s buffer + QPD-OTf (20 µM); green: QPD fluorescence. h Control conditions for experiment reported in d (QPD-OTf 20 µM, 2 h in Ringer’s buffer); green: crystals. Scale bar 20 µM. Source data are provided as Source Data file.