Figure 2. MEL-28 Is an NE Protein during Interphase and Associates with Chromosomes in Mitosis. (A) Immunolocalization of MEL-28 (green), mAb414-reactive nucleoporins (red), and chromatin with Hoechst (blue) showed a dynamic behavior, including NE accumulation (interphase) and kinetochore localization (metaphase) for MEL-28. The scale bar represents 5 um.(B) Immuno-gold localization of MEL-28 in embryos from heterozygous (upper two rows) or homozygous (lower row)
mel-28(
t1684) nematodes revealed an enrichment at NPCs. NE membranes (yellow) and gold particles (red) are highlighted (right column). The scale bar represents 100 nm.(C) Immunolocalization of MEL-28 (green) in embryos expressing GFP-histone H2B (red) indicated that MEL-28 was mostly associated with the chromosome surfaces facing the spindle poles through metaphase and anaphase (three right columns). The scale bar represents 5 um.(D) Colocalization of MEL-28 (red) with the kinetochore component GFP-HIM-10 (green). Microtubules are light blue, and DNA is stained with Hoechst (dark blue). The scale bar represents 10 um.(E) Immunolocalization of MEL-28 (green), microtubules (red), and chromatin (blue) revealed a stable and microtubule-independent kinetochore localization of MEL-28 during metaphase both before (0 s) and after treatment on ice for 30 or 60 s prior to fixation. The scale bar represents 5 um.