FIGURE 2:

Tm1A and Tm1J colocalize in spreading cells but do not colocalize in nonspreading cells. (A–D) Live-cell imaging of S2 cells spreading on ConA with mRFP-actin (red), a marker of the lamellipod, and eGFP-tagged proteins (green): Arp2 (A), Tm1A (B), Tm1J (C), and Tm2A (D). Arp2 localizes to the lamellipod with mRFP-actin, whereas Tm1A and Tm1J both localize to the lamellum and are excluded from the lamellipod. Tm2A shows no significant localization. Right, normalized average fluorescence intensity line scans. (E) Coexpression of eGFP-Tm1J (green) and mCherry-Tm1A (red) in S2 cells on PDL show that in the same cell, Tm1A localizes to the cortex, whereas Tm1J localizes to cytoplasmic ring-like structures. (F–J) Live-cell imaging of interphase S2 cells on PDL with eGFP-tagged proteins: Tm1A, Tm1J, Tm2A, myosin-II, and Diaphanous (Dia). Tm1A (F) and myosin-II (I) colocalize to the cortex (arrowheads), whereas Tm1J (G), Tm2A (H), and Dia (J) localize to cytoplasmic ring-like structures (arrows). mCherry-α-tubulin was coexpressed (not shown) to verify that cells were in interphase. See also Supplemental Figure S2.