Effect of hyper-and hypoactivation of Notch signaling in IL-4-stimulate THP-1

MSCV-GFP-IRES (IRES), MSCV-GFP-Myc-NIC1 (NIC1) and MSCV-GFP-DNMAML (DNMAML) were retrovirally transduced in to THP-1 cell (pCL-Ampho was used as packaging plasmid). GFP positive cells were sorted for selective desired cell. Then IRES, NIC1 and DNMAML overexpressing THP-1 were pretreated with PMA (5ng/ml) for 48 h before stimulation with IL-4 (20 ng/ml) for 3 h. IL-4-stimulated NIC1, IRES or DNMAML overexpressing THP-1

MSCV-GFP-IRES（IRES）、MSCV-GFP-Myc-NIC1（NIC1）与MSCV-GFP-DNMAML（DNMAML）经逆转录病毒转导至THP-1细胞（以pCL-Ampho作为包装质粒）。随后通过分选获取GFP阳性的目的细胞。接着，分别对过表达IRES、NIC1及DNMAML的THP-1细胞采用5ng/ml的佛波酯（PMA）预处理48小时，再以20ng/ml的白细胞介素4（IL-4）刺激3小时。经IL-4刺激的过表达NIC1、IRES或DNMAML的THP-1细胞