Extended Data Fig. 7: Compartment-specific characterization of VCP by SMT and Western-blot.
(a) Composite fluorescence image of Hoechst and Potomac Red channels used to segment nuclear, perinuclear, and cytoplasmic fractions. Scale bar represents 25 μm. (b) Representative image of trajectories within nuclear fraction, perinuclear fraction (c) and cytoplasmic fraction (d). (e) State array distributions comparing NTC to DMSO for each cell compartment. (f) State array distribution for each cell compartment for CB-5083 (left) and FAF2 (right) with dashed regions representing S.D. Each condition is sampling a minimum of 36 FOVs from 6 wells from 2 plate replicates. (g) Western blot quantification of protein levels per cellular compartment following siRNA knockdown of FAF2 at 48 hours post-transfection. (h) and (i) Contribution of well-to-well, FOV-to-FOV, and cell-to-cell biases to 2D jump length, assessed using jump resampling. Variance over sample means as a function of sample size for different resampling procedures. A straight line with slope -1 is the expectation from the law of large numbers; sublinearities are due to residual variances over wells, FOVs, or cells. Number of jumps per well (j), FOV (k), or cell (l) used in these analyses. Heavy dashed line represents median value and light dashed lines represent 1st and 3rd quartiles.
