Phase separation and DAXX redistribution contribute to LANA nuclear body and KSHV genome dynamics during latency and reactivation
Fig 2
LLPS disrupting agent effects on DAXX and ATRX foci.
A. iSLK RFP-LANA cells untreated or treated for 10 min with 3.5% 1,6-Hexanediol were imaged by IF for DAXX (green), RFP-LANA (red) or DAPI (blue). Scale bar = 10 μm. B. iSLK RFP-LANA cells untreated or treated for 10 min with 3.5% 1,6-Hexanediol were imaged by IF for ATRX (green), RFP-LANA (red) or DAPI (blue). Scale bar = 10 μm. C. Same as in panel A, except combined imaging with RFP-LANA (red), DAXX (blue), and ATRX (green). In enlarged images, arrows indicate colocalizations of ATRX and DAXX foci, but lacking LANA. Scale bar = 10 μm. D. Quantification of DAXX (left) and ATRX (right) foci in cells treated as described in panels A and B, respectively. Error bars are s.d., * = p value <0.05, *** = p value <0.001, two-tailed t-test, relative to untreated cells. E. IP-Western analysis of iSLK RFP-LANA cells treated with 3.5% 1,6-Hexanediol or DMSO control for 1 hr. Protein complexes were immunoprecipitated by antibodies to LANA, DAXX, or control IgG, and assayed by Western with antibodies specific for LANA, DAXX, ATRX, or Actin, as indicated. Molecular weight markers are indicated in KDa, and input represents 10% of cell lysis used for each IP.