Figure 5.

LPC blocks diffusion channel formation. Nuclei were assembled for 30 min at 14°C, and the preparation was verified for the absence of diffusion channels, as in Figure 3B, with Alexa488-10 kDa dextran (left panels). The reaction was then supplemented with buffer, LPC, or LPC plus OA, followed by incubation for 90 min at 14°C to assess the affect of LPC on channel formation. The nuclei were assayed for channel formation with Alexa488-10 kDa dextran. A block to channel formation was observed with LPC (asterisk). When LPC and OA were added together, however, diffusion of Alexa488-10 kDa dextran into the nuclei was now observed. Control nuclei assembled at room temperature for 60 min that were the treated with Triton X-100 before the addition of antibodies served as a positive control for the anti-DNA antibody assay and for the dextran quenching assay (bottom panels). Scale bar, 10 μm.