hsp-16.2 transgenes colocalize with pores after HS, and are near pores before. (A) SR-SIM of WT worm embryo immunostained with rabbit anti–LMN-1 (Ce lamin; red) and anti-nucleoporin FG repeat (mAb414; green). See Video 1 for 3D imaging. Below are higher magnifications of an apical and a mid-nuclear view with separated channels. (B–D) Single mid-nucleus focal planes from SR-SIM, showing hsp-16.2 tg1 localization by GFP-lacI signal (white) relative to LMN-1 (red) or NPC (green). Bar, 1 µm. (E) Scoring of hsp-16.2 tg #1 and MosSCI insertion tel V before and after HS (hatched), as in B–D. White, colocalizing with NPC (pore); orange, touching NPC; red, adjacent to lamina. Non-peripheral transgenes are not indicated. The number of hsp-16.2 tg counted, n colocalizing with NPC, n touching NPC, and n adjacent to LMN-1 were as follows. no HS, n = 197, 10, 68, and 35; and after HS, n = 199, 106, 23, and 18. For tel V, numbers were as follows. no HS, n = 215, 45, 46, 44; and after HS (n = 184, 33, 44, 42. The red line is defined as in Fig. 2. Bars: (A, top) 10 µm; (A, bottom) 3 µm; (B–D) 1 µm.