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. Author manuscript; available in PMC: 2024 Jun 6.
Published in final edited form as: Cell Metab. 2023 Apr 25;35(6):1057–1071.e12. doi: 10.1016/j.cmet.2023.04.003

Figure 3. FLVCR1 mediates choline import in mammalian cells.

Figure 3.

a) Schematic for radioactive choline uptake assays using 3H-choline. b) Uptake of [Methyl-3H]-Choline in FLVCR1-knockout HEK293T cells expressing a vector control or FLVCR1 cDNA incubated for indicated timepoint. Data represented as mean ± standard deviation and normalized by seeded cell number; n = 3. Statistical significance was determined by multiple two-sided unpaired t-tests with correction for multiple comparisons using the Holm-Sidak method. t=1 minute p=1.3×10−3; t =2 minutes p=2.3×10−5, t=5 minutes, 15 minutes, and 30 minutes p<1×10-6. c) Uptake of [Methyl-3H]-Choline in FLVCR1-knockout HEK293T cells expressing a vector control or FLVCR1 cDNA incubated with indicated dose for 30 minutes. Data represented as mean ± standard deviation and normalized by seeded cell number; n = 3. Statistical significance was determined by multiple two-sided unpaired t-tests with correction for multiple comparisons using the Holm-Sidak method. 1.34μM p=6×10−6; 2.69μM, 5.38μM, 10.75μM, and 21.5μM p<1×10-6. d) Uptake of 21.5μM [Methyl-3H]-Choline in FLVCR1-knockout HEK293T cells expressing an empty vector (EV) control, SLC44A1, FLVCR1, or SLC5A7 cDNA for 30 minutes. Data represented as mean ± standard deviation and normalized by seeded cell number; n = 3. Statistical significance determined by two-tailed unpaired t-test. e) Phosphocholine abundance in FLVCR1-knockout HeLa cells expressing a vector control or FLVCR1 cDNA after incubation with 1mM phosphocholine or 1mM choline for 24 hours. Bars represent mean ± standard deviation; n = 3. Statistical significance determined by two-tailed unpaired t-test.