Figure 5.
Lfc specifies MLC localization at the cell periphery. (a) An MLC-GFP–expressing DC migrating under agarose along a soluble CCL19 gradient. Central (orange box) and peripheral (periph.; purple box) MLC accumulation is outlined. The blue line indicates the position of the nucleus. The red line outlines cell shape. Scale bar, 10 µm. (b and c) MLC accumulation during migration under agarose in wild type (b) or Lfc−/− (c) cells. Scale bar, 10 µm. Middle panels indicate cell shapes over time. Right panels indicate mean MLC fluorescence distribution along the anterior-posterior polarization axis (dashed line) in 80-s intervals. Arrowheads indicate peripheral (purple) and central (orange) MLC accumulation. (d) Localization of MLC accumulation during directed migration of Lfc+/+ (red) and Lfc−/− (blue) DCs. To account for differences in cell length, the distance between cell center and MLC accumulation was normalized to cell length. Graph shows the distance over time of n = 7 migratory cells per condition ± SD. See also Video 8. (e) Left panel: Localization of endogenous phospho-MLC(S19) (pMLC) in fixed migratory DCs. The blue line indicates the position of the nucleus. The red line outlines cell shape. Right panel indicates the position of MLC accumulation relative to cell length of n = 16 cells per condition from N = 4 experiments. Boxes extend from 25th to 75th percentiles. Whiskers span minimum to maximum values. Annotation above columns indicates results of unpaired Student’s t test; ****, P ≤ 0.0001. Scale bar, 10 µm.

Lfc specifies MLC localization at the cell periphery. (a) An MLC-GFP–expressing DC migrating under agarose along a soluble CCL19 gradient. Central (orange box) and peripheral (periph.; purple box) MLC accumulation is outlined. The blue line indicates the position of the nucleus. The red line outlines cell shape. Scale bar, 10 µm. (b and c) MLC accumulation during migration under agarose in wild type (b) or Lfc−/− (c) cells. Scale bar, 10 µm. Middle panels indicate cell shapes over time. Right panels indicate mean MLC fluorescence distribution along the anterior-posterior polarization axis (dashed line) in 80-s intervals. Arrowheads indicate peripheral (purple) and central (orange) MLC accumulation. (d) Localization of MLC accumulation during directed migration of Lfc+/+ (red) and Lfc−/− (blue) DCs. To account for differences in cell length, the distance between cell center and MLC accumulation was normalized to cell length. Graph shows the distance over time of n = 7 migratory cells per condition ± SD. See also Video 8. (e) Left panel: Localization of endogenous phospho-MLC(S19) (pMLC) in fixed migratory DCs. The blue line indicates the position of the nucleus. The red line outlines cell shape. Right panel indicates the position of MLC accumulation relative to cell length of n = 16 cells per condition from N = 4 experiments. Boxes extend from 25th to 75th percentiles. Whiskers span minimum to maximum values. Annotation above columns indicates results of unpaired Student’s t test; ****, P ≤ 0.0001. Scale bar, 10 µm.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal