Establishment of the LILRA3-overexpressing U937 cell line
The open reading frame (ORF) of LILRA3 was amplified from U937 cells
using qRT-PCR and cloned into the expression vector pWSLV-02 (carrying
enhanced green fluorescent protein (EGFP) as a reporter gene)
(Viewsolid, Beijing, China) to generate the plasmid pWSLV-02-LILRA3. For
lentivirus production, 293T cells were co-transfected with the
pWSLV-02-LILRA3 plasmid, which encodes the intact sequence of LILRA3
cDNA, or the empty vector pWSLV-02 as a control using Lenti-Pac HIV
Expression Packaging Kit (GeneCopoeia, Rockville, USA). Virus was
collected at 24 hours and 48 hours after transfection. To prepare the
LILRA3-overexpressing cell line, U937 cells were infected with
lentivirus for 72 hours, and FCM was then used to select cells with
stable LILRA3 lentivirus integration.