Ha tag antibody

The antibody used for western blot analysis are listed as follows: JMJD2D antibody (Abcam, Cat# ab93694, 1:1000); β-actin antibody (Sigma Aldrich, Cat# A5441, 1:1000); EpCAM antibody (Cell Signaling Technology, Cat# 93790, 1:1000); β-catenin (Cell Signaling Technology, Cat# 8480, 1:1000); TCF4 (Cell Signaling Technology, Cat# 2569, 1:1000); Flag-tag antibody (Sigma, Cat# F1804, 1:1000); Myc-tag antibody (ABclonal, Cat# AE070, 1:1000); H3K9me3 antibody (Abcam, Cat# ab8898, 1:1000); Sox9 (Abcam, Cat# ab182579, 1:1000); Notch1 (Cell Signaling Technology, Cat# 3608, 1:1000); HA-tag antibody (Thermo Fisher Scientific, Cat# 26183, 1:1000). The JMJD2D inhibitor 5-c-8HQ (HY-12304) was purchased from MCE. Lipofectamine 2000 (Cat #11668019) was purchased from Invitrogen.

At 48 h post-transfection, cells were dissociated by TrypLE^TM Select (Invitrogen, USA) and washed once with cold PBS by centrifugation for 5 min at 1,000 × g and 4 °C. The supernatant was removed by aspiration, and the cells were resuspended in 500 μl of FACS buffer (PBS supplemented with 0.5% BSA and 0.1% sodium azide). The cells were analysed using a Cytomics FC 500 MPL flow cytometry system (Beckman Coulter, USA). The data were analysed using MXP software (Beckman Coulter, USA). To select MARC-145 cells stably expressing Cas13b after infection with lentivirus, harvested cells were fixed in 4% paraformaldehyde and permeabilized with ice-cold 90% methanol. After washing, cells were stained with a 1:50 dilution of an HA tag antibody (Thermo Fisher). For secondary staining, a 1:100 dilution of goat anti-mouse IgG1-FITC was added to cells and incubated in the dark. For each measurement, at least 10,000 events were gated.