Fig. 2.

Phosphorylation of KAP1-S824 is necessary for AAV2 transcription and replication. (A) p-KAP1-S824 in 293T cells infected with 10–10,000 genome-containing particles (gcp) per cell of either AAV2 or rAAV2 in the presence of Ad5. (B) ChIP-qPCR performed as described in Fig. 1 D and E in 293T cells infected with AAV2 (100 IU per cell) with or without Ad5; n = 1. (C and D) AAV2 genome replication in 293T cells overexpressing KAP1^WT (C) or KAP1^S824D (D). Values are reported as mean ± SEM, n = 4. (E and F) AAV2 replication in control cells and KAP1-depleted cells reconstituted with KAP1^WT, KAP1^S824D, or KAP1^S824A or treated with an empty vector (EV) control. (E) Viral genome replication. Values are reported as mean ± SEM, n = 4. (F) AAV2 capsid (VP) and KAP1 protein levels. (G) AAV2 transcription in control cells and KAP1-depleted cells complemented with KAP1^WT, KAP1^S824D, or KAP1^S824A or treated with an empty vector (EV) control 16 h after infection with AAV2 (1 IU per cell) alone. Values are reported as mean ± SEM, n = 4. Statistical significance was determined by unpaired t test, *P < 0.05, **P < 0.01, ***P < 0.001.