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. 2023 Mar 3;299(4):104578. doi: 10.1016/j.jbc.2023.104578

Figure 1.

Figure 1

CRISPR KO of CEPT1 and CHPT1 in U2OS cells.A, schematic of the CDP-choline (Cho) and CDP-ethanolamine (Etn) pathways. B, lysates of WT U2OS and two CEPT1 KO cell lines were immunoblotted with antibodies against CEPT1 and actin. A nonspecific band is indicated by the asterisk. C, lysates of CEPT1-KO cells transiently transfected with empty vector (Mock) or pT7-CEPT were immunoblotted with the antibodies against CEPT1, T7, and actin. D, 120 bp deletion in exon 1 of CHPT1 introduced by two gRNAs was screened by PCR amplification and agarose gel electrophoresis. CHPT1-KO1-3 are homozygous for the CRISPR deletion in exon 1. E and F, mRNA for CDP-choline pathway enzymes in CEPT1-KO (panel E) and CHPT1-KO cells (panel F) was quantified by qPCR analysis and expressed relative to U2OS cells. Results are the mean and SD of three biological replicates. CEPT1, choline/ethanolamine phosphotransferase 1; CHPT1, choline phosphotransferase 1.