hLGR5

Fig.1 Detection of LGR5 expression in frontal lobe by RT-PCR. Frontal lobe RNA was extracted from 8-year-old male WT C57BL/6 and homozygous hLGR5 knockin mice (n=3 in each group), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mRNA primers. Mouse Lgr5 mRNA (153 bp) was detectable only in wild-type C57BL/6 mice. Human LGR5 mRNA (142 bp) was detectable only in homozygous hLGR5 knockin mice but not in wild-type mice. 

Abbr. M, marker; HO, homozygous; WT, wild type.

Fig.2 Detection of LGR5 expression in frontal lobe by qPCR. Frontal lobe RNA were extracted from 8-year-old male WT C57BL/6 and homozygous hLGR5 knockin mice (n=3 in each group), then cDNA libraries were synthesized by reverse transcription, followed by qPCR with h/m Lgr5 and mouse Gapdh primers. Relative expression represents the Lgr5 mRNA level relative to its average expression in WT mice. Lgr5 mRNA was expressed similarly in wild-type mice and homozygous hLGR5 knockin mice by detecting with a h/m Lgr5-shared primer.

Fig.3 Detection of LGR5 expression in hLGR5 mice by WB. The frontal lobe, lung, colon and liver tissue lysates were collected from 9-week-old male wild-type C57BL/6 mice and homozygous hLGR5 mice, and then analyzed by western blot with anti-LGR5 antibody. LGR5 was clearly detected in the lung, colon, and liver tissues, with low expression in the frontal lobe tissues of both WT C57BL/6 and HO hLGR5 mice, as the antibody was cross-reactive with both human and mouse LGR5.

Abbr. HO, homozygous; WT, wild type; PC, positive control, HepG2 cells.

Fig.4 Detection of LGR5 expression in hLGR5 mice by WB. The stomach tissue lysates were collected from 9-week-old male WT C57BL/6 mice and homozygous hLGR5 mice, then analyzed by western blot with anti-LGR5 antibody. LGR5 was detectable in stomach from both WT C57BL/6 and HO hLGR5 mice , as the antibody was cross-reactived between human and mouse LGR5. 

Abbr. HO, homozygous; WT, wild type; PC, positive control, HepG2 cells.