Rabbit anti utx

The inserts of plasmids pcs2-UTX and pcs2-ZSCAN4D used in this study were PCR-amplified from the cDNA of NIH-3T3 cells and 2-cell mouse embryos, respectively. The PCR products were cloned into the Xho I and Xba I (Thermo, USA) sites downstream of the Myc ectopic tag. The antibodies covered were rabbit anti-UTX (Millipore, USA, for IF and WB), rabbit anti-H3K27me3 (Abcam, USA, for IF), mouse anti-Myc (Santa Cruz, USA, for IF), rabbit anti-ZSCAN4 (Millipore, for IF), rabbit anti-CDX2 (Abcam, for IF), mouse anti-NANOG (Abcam, for IF), rabbit α-tubulin (Proteintech, USA, for WB), Alexa Fluor 594 and Alexa Fluor 488 (Thermo, for IF), anti-UTX (KDM6A) (Abcam, for ChIP), anti-IgG (Abcam, for ChIP).

Immunostaining was performed on paraformaldehyde (PFA)-fixed paraffin-embedded adult pancreas tissues as previously described [69 (link)]. Primary antibodies included rabbit anti-Mll1 (1/250; Bethyl Laboratories), rabbit anti-Mll3 (1/250; Abcam), rabbit anti-Wdr5 (1/2500; Bethyl Laboratories), rabbit anti-Dpy30 (1/500; Abcam), rabbit anti-Jmjd3 (1/50; Millipore), rabbit anti-Utx (1/2000; Millipore), and guinea pig anti-insulin (1/1000; Abcam). Fluorescently conjugated secondary antibodies included donkey anti-rabbit Alexa 488 (1/1000; Molecular Probes) and goat anti-guinea pig Alexa 546 (1/500; Molecular Probes). Sections were imaged with an SP8 confocal microscope (Leica).