Disruption of AKA95-RII Anchoring or cAMP/PKA Inhibition Does Not Inhibit Mitotic Chromosome Condensation
| Inhibitor . | Mitotic index . |
|---|---|
| (Percent) | |
| None (∼25 pl FITC-dextran) | 88 ± 8 |
| α-AKAP95 (polyclonal; ∼2 pg) | 17 ± 7 |
| α-AKAP95 (∼2 pg) + GST-AKAP95Δ1-386 (250 pg) | 80 ± 6 |
| Ht31 (50 nM) | 79 ± 8 |
| Ht31-P (50 nM) | 82 ± 6 |
| PKI (10 nM) | 85 ± 5 |
| Rp-8-Br-cAMPS (10 μM) | 90 ± 4 |
| Inhibitor . | Mitotic index . |
|---|---|
| (Percent) | |
| None (∼25 pl FITC-dextran) | 88 ± 8 |
| α-AKAP95 (polyclonal; ∼2 pg) | 17 ± 7 |
| α-AKAP95 (∼2 pg) + GST-AKAP95Δ1-386 (250 pg) | 80 ± 6 |
| Ht31 (50 nM) | 79 ± 8 |
| Ht31-P (50 nM) | 82 ± 6 |
| PKI (10 nM) | 85 ± 5 |
| Rp-8-Br-cAMPS (10 μM) | 90 ± 4 |
Nuclei of HeLa cells synchronized in interphase were injected with either 2–5 pg affinity-purified polyclonal anti–AKAP95 antibodies, 2–5 pg anti–AKAP95 antibodies with 250 pg GST-AKAP95Δ1-386 peptide, 50 nM RII-anchoring inhibitor peptide Ht31, 50 nM control Ht31-P peptide, 10 nM PKI, or 10 μM cAMP antagonist Rp-8-Br-cAMPS. Injection volumes were 25–50 pl. Control nuclei were injected with 25–50 pl FITC-dextran only. Cells were released from the thymidine block and synchronized with 1 μM nocodazole to induce M phase arrest. After 17 h in nocodazole, mitotic indexes were determined. Data from 30–50 injected cells per treatment are reported. (Similar results were obtained regardless of the site of injection within the cells; data not shown).