Extended Data Fig. 5: Mass spectrometry imaging reveals metabolic alterations following Slc7a5 deletion.
From: The amino acid transporter SLC7A5 is required for efficient growth of KRAS-mutant colorectal cancer
a, Schematic of workflow for LC-MS and mass spectrometry imaging (MSI) of intestinal tissue. b, H&E and ion intensities of glutamine, glutamate and alanine in Apcfl/fl KrasG12D/+ Slc7a5+/+ and Apcfl/fl KrasG12D/+ Slc7a5fl/fl small intestinal tissue, using MSI. Representative ion images from three biologically independent mice for each genotype. c, Table showing ion intensities from b. d, Abundance of amino acids (metabolite peak area/µg of protein) in Apcfl/fl KrasG12D/+ Slc7a5+/+ and Apcfl/fl KrasG12D/+ Slc7a5fl/fl small intestinal tissues using LC-MS. Heatmap shows mean relative abundance (n = 4 biological replicates per group). Asterisks denote P, 0.001 (Met), 0.004 (His), 0.01 (Gly), 0.02 (Pro), 0.02 (Asp), 0.003 (Lys), 0.0001 (Ser), 0.0005 (Thr), 0.000006 (Arg), 0.002 (Gln), using two-way ANOVA with Fisher’s test. e, H&E and autoradiogram of O-(2-18F-fluoroethyl)-L-tyrosine (18F-FET) uptake and distribution in Apcfl/fl KrasG12D/+ Slc7a5+/+ and Apcfl/fl KrasG12D/+ Slc7a5fl/fl mice. Representative images from four biologically independent mice for each genotype. f, H&E and ion intensities of leucine, 13C6-leucine, KIC and KIC M+6 in WT, KrasG12D/+, Apcfl/fl, Apcfl/fl KrasG12D/+ Slc7a5+/+ and Apcfl/fl KrasG12D/+ Slc7a5fl/fl small intestinal tissue, generated using MSI. Representative ion images from four individual mice for each genotype. g, Ratio of ion abundances of KIC M+6 and 13C6-leucine. Plot shows mean ratios ± s.e.m. h, Ion abundance of citrate M+2 (detected with MSI) and peak area of acetyl-CoA M+2 (detected with LC-MS). Plots show their respective mean ± s.e.m. In g, h WT, n =5, Apcfl/fl, Apcfl/fl KrasG12D/+, KrasG12D/+, and Apcfl/fl KrasG12D/+ Slc7a5fl//fl, n = 4; per group. P using one-way ANOVA followed by Tukey’s test. In b, f selected ion images representing the accurate mass distribution of indicated metabolites are shown. Ion intensities were scaled individually for each metabolite and m/z ratios indicated.
