Nuclear translocation is accompanied with forward movements of microtubule tracks. (A) Representative time-lapse sequence of a cultured CGC transfected with mEos2-DCX. The nucleus was labeled with SiR-DNA. At 2 min, a strong 405-nm laser is applied at a point (denoted by a blue circle) in the proximal region of the leading process for green-to-red photoconversion of mEos2-DCX. (B) Kymograph (left) and intensity distribution (right) of fluorescent signals of the cell in A show synchronous forward movement of the nucleus (blue) and photoconverted MTs (red). (C and D) Representative time-lapse sequences of Nesprin-2 mutant CGCs (C) or WT CGCs overexpressing HA-KASH (D). Photoconversion is induced at 2 min at the points demoted by blue circles. (E) Quantification of MT forward moving speed. Unpaired t test is used to compare to WT. (F) Quantification of changes in Nuclear-MT (NM) distance in migrating CGCs. The change of NM distance over 3–6 min is normalized to the initial NM distance. Unpaired t test with Welch’s correction to Ctrl samples. n = 22 (WT), 12 (Nesp2 mutant), and 19 (WT + HA-KASH) cells from two to six independent experiments per group. (G) Model of nuclear translocation mechanism. Forward movements of nucleus are generated by the combination of the persistent Nesprin-2-mediated bidirectional transport and MT forward movements in CGCs. Bars show mean ± SEM. Scale bars, 5 μm. Related to Video 10.