Figure 3.

NM2 anterograde movements independent of actin dynamics

(A) Cartoon of C-terminal HaloTag on an NM2 monomer, filament, and imaged with high-resolution microscopy. (B and D) CAD cell expressing NM2A-Halo imaged before (B) or after (D) treatment with jasplakinolide/latrunculin (JL) cocktail. Purple shaded boxes indicate ROIs for (C) and (E). (C and E) Kymographs shown without (left) and with (right) retrograde (blue dotted lines) and anterograde (orange dotted lines) movements indicated. (F) Rose plot of individual retrograde (blue) and anterograde (orange) tracks detected in a single cell before and after JL treatment. (G) Example lamellar regions of NM2A-Halo used for quantification of leading edge intensity (orange outline) in (H) before and 45 s after JL treatment. (H) Ratio of NM2A-Halo intensity within 1 μm of the leading edge before and 45 s after JL treatment. Imaging was performed on Zeiss 880 Aiscan.