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. 2023 Dec 8;5(12):2148–2168. doi: 10.1038/s42255-023-00936-2

Fig. 5. SLC6A14 LeuT domain mediates serine transport in colorectal cancer cells.

Fig. 5

a, Amino acid uptake or release levels (pmol per cell) over 24 h of HCT116 cells NTC (siRNA control) or silenced for SLC6A14. Control data (NTC) from the same experiment are also shown in Figs. 3i and 4h. b, Growth curves of HCT116 cells expressing EVEGFPN1 (empty vector, control) or GFP-tagged SLC6A14. Values are mean ± s.d. from n = 3 biological replicates and representative of three independent experiments. Statistical significance assessed by two-tailed Welch’s t-test at t = 87 h. c, AUC from uptake curves of SLC6A14-silenced HCT116 cells labeled with a pool of labeled amino acids as indicated for 1, 3 and 6 min. Values are mean ± s.d. and relative to NTC siRNA control from n = 3 biological replicates. Statistical significance was assessed with two-tailed one-sample t-test on natural log-transformed values. d, [13C3,15N]l-serine consumption in HCT116 SLC6A14-depleted cells overexpressing either EVEGFPN1 (EV, control) or SLC6A14EGFPN1. Values are mean ± s.d. from n = 3 biological replicates. Statistical significance assessed by two-tailed Welch’s t-test. e, Structure of the human SLC6A14 protein as predicted by AlphaFold2. f, Sequence alignment of different LeuT-containing transporters to predict equivalent residues to V104 of the LeuT transporter. g, Sequence alignment of SLC6A14 protein between different organisms. h, Cartoon specifying mutagenesis strategy on SLC6A14 protein. i, [13C3,15N]l-serine consumption in HCT116 cells overexpressing EGFPN1 (EV control) or GFP-tagged SLC6A14 WT, -V128W, -V128G and -V128E. Values are mean ± s.d. from n = 3 biological replicates. Statistical significance was assessed using ordinary ANOVA and Dunnett’s multiple comparisons test.