Propidium iodide p3566

Cells (1 × 10⁵) were collected from uninfected OECs, HCMV-infected OECs and CTO cells. Cells were fixed, permeabilized, and stained as previously reported [19 (link)]. Cytofluorometric analysis was achieved using a BD LSRFortessa X-20 (BD Biosciences) flow cytometer. FACSDiva software (BD Biosciences) was used for data collection and analysis. The antibodies used are provided in Supplementary Table 2. For cell cycle analysis, uninfected OECs or CTO cells were washed in 1X PBS, fixed in 70% ethanol, and resuspended in 50 µg/ml propidium iodide (P3566, life technologies, Eugene, USA) with 0.1 mg/ml RNase (R4642, Sigma-Aldrich, Saint-Louis, MO, USA), then incubated at 37 °C for 30 min as described previously [17 (link)].

Rabbit antibodies against TRIM28 (4124S, 4123S), CTCF (3418S), OCT4A (2890S), SOX2 (23064S), NANOG (3580S), KLF4 (12173S), SUMO2 (4971S), and mouse antibodies against TRA-1-60 (4746T), TRA-1-81(4745T), SSEA-4 (4755T), and SMA (D4K9N) were from Cell Signalling Technology. Anti-SALL4 (ab29112), anti-TRIM24 (ab70560), anti-NOP58 (ab155556), anti-NESTIN (ab196908) were from Abcam. Mouse antibody against α-Tubulin was from Bethyl Laboratories, mouse anti-LaminA/C antibody was from Sigma Aldrich (SAB4200236), rabbit anti-mCherry (PA5-34974), rabbit anti-TRIM33 (PA5-82152) were from Invitrogen, and mouse anti-HIS (34650) was from Qiagen. Rabbit anti-CYTOKERATIN17 was a gift from R. Hickerson (University of Dundee). Sheep antibodies against SUMO1, SUMO2 (27 (link)) were generated in-house. Secondary antibodies conjugated with HRP and Alexa fluorophores, Cy3 Cellmask (C10045), Rhodamine Phalloidin (R415) were from Sigma and Invitrogen, respectively. ML792 (1644342-14-2), MG132 (474787), and N-ethylmaleimide (E3876) were from Sigma Aldrich. Protease Inhibitor cocktail (11836170001) was from Roche. Propidium iodide (P3566) and DAPI (D1306) were from Life Technologies.

Domatinostat (4SC-202; S7555) was purchased from Selleck Chemicals (Houston, TX, USA). Domatinostat was dissolved in DMSO to prepare a 10 mM stock solution. Propidium iodide (P3566) and Hoechst33342 (H3570) solutions were purchased from Thermo Fisher Scientific Inc. (Waltham, MA, USA). An antibody against FOXM1 (ab207298) was purchased from Abcam plc (Cambridge, UK). Antibodies against Survivin (#2808), GAPDH (#5174), cleaved PARP (#9541), and cleaved caspase 3 (#9661) were purchased from Cell Signaling Technology Inc. (Beverly, MA, USA). Horseradish peroxidase (HRP)-conjugated anti-rabbit IgG and anti-mouse IgG secondary antibodies were purchased from Jackson ImmunoResearch (West Grove, PA, USA).