Journal: bioRxiv
Article Title: The targeted cytosolic degradation of class I histone deacetylases is essential for efficient alphaherpesvirus replication
doi: 10.64898/2026.01.02.697337
Figure Lengend Snippet: (A) Immunoblotting analysis of the indicated proteins in HeLa cells either mock-infected or infected with HSV-1 (MOI = 1), treated with vehicle, 3-MA (10 μM), MG-132 (10 μM), or chloroquine (10 μM) for the indicated times. (B) Ubiquitination assay of HDAC1 in HeLa cells either mock-infected or infected with HSV-1 (MOI = 1) at 24 hpi. (C) Ubiquitination assay of FLAG-HDAC1 in HeLa cells transfected with HA-ubiquitin variants (WT, K48, K63), and either mock-infected or infected with HSV-1 (MOI = 1) at 24 hpi. (D) Schematic representation of HDAC1 deletion mutants. (E-I) Ubiquitination assays of FLAG-HDAC1 mutant variants in HeLa cells either mock-infected or infected with HSV-1 (MOI = 1) at 24 hpi. (J) Ubiquitination assays of FLAG-HDAC2 variants (WT and K75R) in HeLa cells either mock-infected or infected with HSV-1 (MOI = 1) at 24 hpi.
Article Snippet: Antibodies against CHK1 (25887-1-AP), CHK2 (13954-1-AP), RAD51 (14961-1-AP), β-actin (66009-1-lg), P53 (10442-1-AP), HDAC1 (10197-1-AP), HDAC2 (12922-3-AP), HDAC4 (17449-1-AP), HDAC6 (12834-1-AP), HDAC11 (67949-1-Ig), and EGFP (50430-2-AP) were purchased from Proteintech; antibodies against p-P53 (9286), p-ATM (13050), ATM (2873), ATR (13934), p-ATR (2853), p-CHK1 (12302), p-CHK2 (2197), γ-H2AX (80312), H3 (4499), H4K8ac (2594), H4K12ac (13944), H4 (13919), H2AX (7631), H3K9ac (4658), H3K27ac (8173), UB (20326), and MDM2 (86934) were purchased from Cell Signaling Technology; H3K56ac antibody (07-677) was purchased from Millipore; ICP4 antibody (ab6514) was purchased from Abcam; ICP0 antibody (sc-53070) was purchased from Santa Cruz Biotechnology; FLAG antibody (F7425) was purchased from Sigma-Aldrich; HA antibody (A00169) was purchased from GenScript.
Techniques: Western Blot, Infection, Ubiquitin Proteomics, Transfection, Mutagenesis