human slc7a5 (Addgene inc)
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Addgene inc
human slc7a5
Human Slc7a5, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 169 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+slc7a5/pm39756034-230-36-34?v=Addgene+inc
Average 94 stars, based on 169 article reviews
Human Slc7a5, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 169 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+slc7a5/pm39756034-230-36-34?v=Addgene+inc
Average 94 stars, based on 169 article reviews
human slc7a5 - by Bioz Stars,
2026-08
94/100 stars
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1) Product Images from "SLC7A5 is required for cancer cell growth under arginine-limited conditions."
Article Title: SLC7A5 is required for cancer cell growth under arginine-limited conditions.
Journal: Cell reports
doi: 10.1016/j.celrep.2024.115130
Figure Legend Snippet: Figure 1. A functional genomics screen identifies SLC7A5 as required for growth on citrulline (A) Schematic of citrulline synthesis into arginine. (B) Concentrations of arginine and citrulline in mouse serum (mean ± SD, n = 5). (C) Rates of appearance of arginine and citrulline as determined by steady-state labeling fraction upon defined infusion (mean ± SD, n = 5). (D) Media conditions used for the CRISPR screen. (E) Growth of A375mASS1-OE cells in Low Arg, High Arg, and Low Arg minus Cit media. Data are expressed in terms of relative growth, where the readings on day 0 are normalized to 1 (mean ± SD, n = 4). (F) Schematic of the CRISPR-based screen to identify metabolic genes required for growth under low-arginine conditions. (G) Gene scores from cells grown under Low Arg vs. High Arg. SLC7A5, the top hit, is highlighted in red. SLC7A1 (arginine transporter) and ASL (arginosuccinate lyase) are also highlighted. The red line is the equation y = x passing though (0,0) to highlight differential essentiality. (H) Top 25 genes scoring as selectively essential in Low Arg vs. High Arg. Genes linked to glycosylation are shown in blue, the urea cycle in red, transport in purple, reactive oxygen species (ROS) metabolism in orange, and other genes in green.
Techniques Used: Functional Assay, Labeling, CRISPR, Glycoproteomics
![Figure 2. SLC7A5 is required for citrulline uptake, metabolism, and growth in arginine-free media (A) Immunoblot showing SLC7A5 and ASS1 expression in two SLC7A5 KO clones in the A375mASS1-OE (melanoma) cell line. WT, SLC7A5 WT. b-Actin was used as a loading control. (B) Growth of A375mASS1-OE and SLC7A5 KO cells in the indicated media. The base medium was RPMI without arginine supplemented with 110 mM arginine or citrulline as indicated. Media were refreshed on day 2 (mean ± SD, n = 4). (C) Immunoblot showing SLC7A5 expression from two SLC7A5 KO clones in the MCF7 (breast cancer) cell line. b-Actin was used as a loading control. (D) Growth of MCF7 SLC7A5 WT and KO cells in the indicated media. Media were refreshed on day 2. (mean ± SD, n = 4). (E) Citrulline consumption from the media over a 48-h period; A375mASS1-OE and SLC7A5 KO. Media: RPMI + 110 mM Arg + 110 mM Cit (mean ± SD, n = 4). **p < 0.01, unpaired two-tailed t test. (F) Citrulline consumption from the media with SLC7A5 WT and KO over a 48-h period; MCF7 WT and SLC7A5 KO. Media: RPMI + 110 mM Arg + 110 mM Cit (mean ± SD, n = 4). ***p < 0.001, unpaired two-tailed t test. (G) Uptake of [1-13C]-citrulline over a 15-min period in A375m WT and SLC7A5 KO cells. Media: RPMI + 110 mM Arg + 40 mM Cit (mean ± SD, n = 3). ***p < 0.001, unpaired two-tailed t test. (H) Ion count of unlabeled (M+0) and labeled (M+1) argininosuccinate. Cells were plated in RPMI + 110 mM [1-13C]-citrulline for 16 h, and then metabolites were extracted and analyzed using LC-MS (mean ± SD, n = 4).](https://pub-med-unpaywalled-images-cdn.bioz.com/pub_med_ids_ending_with_6034/pm39756034/pm39756034__page5_image1.jpg "Figure 2. SLC7A5 is required for citrulline uptake, metabolism, and growth ...")
Figure Legend Snippet: Figure 2. SLC7A5 is required for citrulline uptake, metabolism, and growth in arginine-free media (A) Immunoblot showing SLC7A5 and ASS1 expression in two SLC7A5 KO clones in the A375mASS1-OE (melanoma) cell line. WT, SLC7A5 WT. b-Actin was used as a loading control. (B) Growth of A375mASS1-OE and SLC7A5 KO cells in the indicated media. The base medium was RPMI without arginine supplemented with 110 mM arginine or citrulline as indicated. Media were refreshed on day 2 (mean ± SD, n = 4). (C) Immunoblot showing SLC7A5 expression from two SLC7A5 KO clones in the MCF7 (breast cancer) cell line. b-Actin was used as a loading control. (D) Growth of MCF7 SLC7A5 WT and KO cells in the indicated media. Media were refreshed on day 2. (mean ± SD, n = 4). (E) Citrulline consumption from the media over a 48-h period; A375mASS1-OE and SLC7A5 KO. Media: RPMI + 110 mM Arg + 110 mM Cit (mean ± SD, n = 4). **p < 0.01, unpaired two-tailed t test. (F) Citrulline consumption from the media with SLC7A5 WT and KO over a 48-h period; MCF7 WT and SLC7A5 KO. Media: RPMI + 110 mM Arg + 110 mM Cit (mean ± SD, n = 4). ***p < 0.001, unpaired two-tailed t test. (G) Uptake of [1-13C]-citrulline over a 15-min period in A375m WT and SLC7A5 KO cells. Media: RPMI + 110 mM Arg + 40 mM Cit (mean ± SD, n = 3). ***p < 0.001, unpaired two-tailed t test. (H) Ion count of unlabeled (M+0) and labeled (M+1) argininosuccinate. Cells were plated in RPMI + 110 mM [1-13C]-citrulline for 16 h, and then metabolites were extracted and analyzed using LC-MS (mean ± SD, n = 4).
Techniques Used: Western Blot, Expressing, Clone Assay, Control, Two Tailed Test, Labeling, Liquid Chromatography with Mass Spectroscopy
Figure Legend Snippet: Figure 4. SLC7A5 and ASS1 are upregu- lated in response to arginine starvation (A) Relative transcript levels of ATF4, ASS1, SLC7A5, and SLC7A1 in A375m WT cells cultured in the indicated media as determined by RT- qPCR. Ct values were normalized to RPS2 (mean ± SD, n = 4). **p < 0.01, ***p < 0.001, ****p < 0.0001; 2-way ANOVA and multiple com- parisons were corrected using the Holm-Sidak method. (B) Immunoblots of SLC7A5 and ASS1 from A375m, HCT116, and LN229 WT cells plated for 48 h in the indicated media. (C) Relative amino acid levels of cells plated for 24 h in either RPMI + 110 mM Cit (Cit, red), RPMI + 110 mM Arg (Arg, blue), RPMI + 110 mM Cit + 1 mM GCN2iB (Cit + GCN2iB, gray) or RPMI + 110 mM Arg + 1 mM GCN2iB (Arg + GCN2iB, gold) (mean ± SD, n = 3–4). ns, not significant; **p < 0.01, ***p < 0.001, ****p < 0.0001, multiple comparisons- corrected 2-way ANOVA. (D) Immunoblot of SLC7A5 and ATF4 in A375m WT cells plated for 48 h with and without GCN2iB (1 mM) in the indicated media containing combi- nations of RPMI + 110 mM Cit or RPMI + 110 mM Arg. (E) Correlation between transcript levels of SLC7A5 versus ASS1 in breast cancer (BRCA) tumor transcript data taken from TCGA. The gray area around the linear regression line indicates the 95% confidence interval. (F) Correlation between transcript levels of SLC7A5 versus ASS1 in the luminal B (LumB) subtype of BRCA tumor transcript data taken from TCGA. The gray area around the linear regression line indicates the 95% confidence interval.
Techniques Used: Cell Culture, Quantitative RT-PCR, Western Blot
Figure Legend Snippet: Figure 5. A small-molecule inhibitor of SLC7A5 sensitizes cells to arginine deprivation (A) Growth of cancer cell lines cultured in RPMI + Arg, RPMI + Cit, with and without 1 mM JPH203. Media were refreshed on day 2 (mean ± SD, n = 4). ***p > 0.001, ****p < 0.0001, 2-way ANOVA. (B) Rescue of growth in the Cit + JPH203 condition with either 1 mM citrulline or 110 mM arginine. Media were refreshed on day 2 (mean ± SD, n = 4). ****p < 0.0001, unpaired two-tailed t test.
Techniques Used: Cell Culture, Two Tailed Test
![Figure 6. SLC7A5 regulates citrulline metabolism in an in vivo xenograft model (A) Immunoblot of SLC7A5 in tumors originating from A375m xenografts. b-Actin was used as a loading control. (B) Growth of bilateral A375m WT and SLC7A5 KO xenograft tumors. Mice were fed a high-protein chow diet (mean ± SEM, n = 10). **p < 0.01, paired two- tailed t test. (C) Labeling fraction of [1-13C]-citrulline in the serum of mice 1 h after intra- peritoneal injection (mean ± SD, n = 5). (D) Tumor [1-13C]-citrulline uptake normalized to serum citrulline. Mice were injected with 83 mM (0.07 g/kg) of [1-13C]-citrulline (mean ± SEM, n = 5). *p < 0.05, paired two-tailed t test.](https://pub-med-unpaywalled-images-cdn.bioz.com/pub_med_ids_ending_with_6034/pm39756034/pm39756034__page11_image1.jpg "Figure 6. SLC7A5 regulates citrulline metabolism in an in vivo xenograft ...")
Figure Legend Snippet: Figure 6. SLC7A5 regulates citrulline metabolism in an in vivo xenograft model (A) Immunoblot of SLC7A5 in tumors originating from A375m xenografts. b-Actin was used as a loading control. (B) Growth of bilateral A375m WT and SLC7A5 KO xenograft tumors. Mice were fed a high-protein chow diet (mean ± SEM, n = 10). **p < 0.01, paired two- tailed t test. (C) Labeling fraction of [1-13C]-citrulline in the serum of mice 1 h after intra- peritoneal injection (mean ± SD, n = 5). (D) Tumor [1-13C]-citrulline uptake normalized to serum citrulline. Mice were injected with 83 mM (0.07 g/kg) of [1-13C]-citrulline (mean ± SEM, n = 5). *p < 0.05, paired two-tailed t test.
Techniques Used: In Vivo, Western Blot, Control, Two Tailed Test, Labeling, Injection