Review

anti slc7a5 (Proteintech)

Proteintech is a verified supplier

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

Proteintech anti slc7a5
Anti Slc7a5, supplied by Proteintech, used in various techniques. Bioz Stars score: 94/100, based on 51 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+slc7a5/SLC7A5+Antibody/pm41792125-375-10-11
Average 94 stars, based on 51 article reviews

anti slc7a5 - by Bioz Stars, 2026-10

94/100 stars

Images

Immunohistochemistry:

Article Title: FBW7/GSK3β mediated degradation of IGF2BP2 inhibits IGF2BP2-SLC7A5 positive feedback loop and radioresistance in lung cancer.
Article Snippet: Anti-IGF2BP2 (#ab128175, Abcam), antim6A (#ab151230, Abcam), anti-rabbit IgG (#3900S, Cell Signaling Technology), and anti-mouse IgG (#61656S, Cell Signaling Technology) were used for RIP and MeRIP assays. .. Anti-IGF2BP2 (#11,601–1-AP, Proteintech, 1:100), anti-FBW7 (#28,424–1-AP, Proteintech, 1:100), and anti-SLC7A5 (#28,670–1-AP, Proteintech, 1:100) were used for immunohistochemistry staining of LUAD tissue microarray. .. Anti-Rad51 (#8875, Cell Signaling Technology) was used for Rad51 foci formation assay.

Staining:

Microarray:

Western Blot:

Article Title: FBW7/GSK3β mediated degradation of IGF2BP2 inhibits IGF2BP2-SLC7A5 positive feedback loop and radioresistance in lung cancer
Article Snippet: .. Anti-IGF2BP2 (#11,601–1-AP, Proteintech, 1:2000), anti-FBW7 (#28,424–1-AP, Proteintech, 1:2000), anti-SLC7A5 (#28,670–1-AP, Proteintech, 1:2000), anti-GSK3β (#22,104–1-AP, Proteintech, 1:1000), anti-H3K4me3 (#A22225, ABclonal, 1:10,000), anti-SETD1A (#A18231, ABclonal, 1:500), anti-mTOR (#66,888–1-Ig, Proteintech, 1:5000), anti-pmTOR-S2448 (#67,778–1-Ig, Proteintech, 1:2000), anti-AKT (#60,203–2-Ig, Proteintech, 1:5000), anti-pAKT-S473 (#66,444–1-Ig, Proteintech, 1:5000), anti-Myc-tag (#60,003–2-Ig, Proteintech, 1:2000), anti-Flag-tag (#66,008–4-Ig, Proteintech, 1:5000), anti-HA-tag (#66,006–2-Ig, Proteintech, 1:10,000), and anti-GAPDH (#60,004–1-Ig, Proteintech, 1:50,000) were used for Western blotting analyses. .. Anti-SETD1A (#61,702, Cell Signaling Technology, 1:100), anti-H3K4me3 (#9751, Cell Signaling Technology, 1:50), and anti-rabbit IgG (#3900S, Cell Signaling Technology, 1:1000) were used for ChIP assay.

Article Title: FBW7/GSK3β mediated degradation of IGF2BP2 inhibits IGF2BP2-SLC7A5 positive feedback loop and radioresistance in lung cancer.
Article Snippet: .. Anti-IGF2BP2 (#11,601–1-AP, Proteintech, 1:2000), antiFBW7 (#28,424–1-AP, Proteintech, 1:2000), anti-SLC7A5 (#28,670–1-AP, Proteintech, 1:2000), anti-GSK3β (#22,104–1-AP, Proteintech, 1:1000), anti-H3K4me3 (#A22225, ABclonal, 1:10,000), anti-SETD1A (#A18231, ABclonal, 1:500), anti-mTOR (#66,888–1-Ig, Proteintech, 1:5000), anti-pmTOR-S2448 (#67,778–1-Ig, Proteintech, 1:2000), anti-AKT (#60,203–2-Ig, Proteintech, 1:5000), anti-pAKT-S473 (#66,444–1-Ig, Proteintech, 1:5000), anti-Myc-tag (#60,003–2-Ig, Proteintech, 1:2000), antiFlag-tag (#66,008–4-Ig, Proteintech, 1:5000), anti-HA-tag (#66,006–2-Ig, Proteintech, 1:10,000), and anti-GAPDH (#60,004–1-Ig, Proteintech, 1:50,000) were used for Western blotting analyses. .. Anti-SETD1A (#61,702, Cell Signaling Technology, 1:100), anti-H3K4me3 (#9751, Cell Signaling Technology, 1:50), and anti-rabbit IgG (#3900S, Cell Signaling Technology, 1:1000) were used for ChIP assay.

Incubation:

Article Title: Targeting deubiquitinase OTUB1 protects vascular smooth muscle cells in atherosclerosis by modulating PDGFRβ.
Article Snippet: Atherosclerosis is a chronic artery disease that causes various types of cardiovascular dysfunction.. Vascular smooth muscle cells (VSMCs), the main components of atherosclerotic plaque, switch from contractile to synthetic phenotypes during atherogenesis.. Ubiquitylation is crucial in regulating VSMC phenotypes in atherosclerosis, and it can be reversely regulated by deubiquitinases.

Cell-Signaling:

Image Search Results

Tcf1 positively regulates Slc7a5 transcription. (A–G) CD4 + T cells were isolated from the spleen and stimulated with anti‐CD3/CD28 antibodies in the presence of MAPK13‐inhibitors (17.1 µ m ) or DMSO for 72 h. The cells were then collected for RNA‐seq, RT‐qPCR, western blotting, and ChIP‐seq. (A) Gene Ontology enrichment analysis showing the top 20 downregulated terms ranked by gene ratio. (B) KEGG pathway enrichment analysis. (C) Volcano plot of RNA‐seq results displaying upregulated and downregulated genes. (D) Relative mRNA expression levels of Tcf7 and Slc7a5 ( n = 3). (E) Protein levels of Slc7a5 ( n = 3). (F) ChIP‐seq tracks of Tcf1 binding at Slc7a5 and Tcf7 loci. Differential binding analysis was performed using MACS2. (G) Dual‐luciferase assay detecting Tcf1 binding to a 100‐bp promoter fragment of Slc7a5 ( n = 3). (H) Predicted Tcf1 binding site in the Slc7a5 promoter fragment identified using JASPAR. (I). Splenic CD4 + T cells stimulated with anti‐CD3/CD28 antibodies for 24 h were infected with lentivirus carrying murine pCDH‐Tcf1 T289E , pCDH‐Tcf1 T289A or pCDH‐Tcf1. CD4 + T cells were collected 48 h post‐infection, and RT‐qPCR was performed to detect the mRNA expression levels of Tcf7 and Slc7a5 ( n = 3). (J) Splenic CD4 + T cells stimulated with anti‐CD3/CD28 antibodies for 24 h were infected with pCDH‐H1‐EF1‐Puro scramble control or murine Tcf1 shRNA retroviruses. RT‐qPCR was performed to detect the mRNA expression levels of Tcf7 and Slc7a5 ( n = 3). ** p < 0.001, *** p < 0.0001, **** p < 0.00001.

Journal: Advanced Science

Article Title: Targeting the Mapk13‐Tcf1‐Slc7a5 Axis via One‐Carbon Metabolic Regulation to Prevent Chronic Allograft Vasculopathy

doi: 10.1002/advs.202520815

Figure Lengend Snippet: Tcf1 positively regulates Slc7a5 transcription. (A–G) CD4 + T cells were isolated from the spleen and stimulated with anti‐CD3/CD28 antibodies in the presence of MAPK13‐inhibitors (17.1 µ m ) or DMSO for 72 h. The cells were then collected for RNA‐seq, RT‐qPCR, western blotting, and ChIP‐seq. (A) Gene Ontology enrichment analysis showing the top 20 downregulated terms ranked by gene ratio. (B) KEGG pathway enrichment analysis. (C) Volcano plot of RNA‐seq results displaying upregulated and downregulated genes. (D) Relative mRNA expression levels of Tcf7 and Slc7a5 ( n = 3). (E) Protein levels of Slc7a5 ( n = 3). (F) ChIP‐seq tracks of Tcf1 binding at Slc7a5 and Tcf7 loci. Differential binding analysis was performed using MACS2. (G) Dual‐luciferase assay detecting Tcf1 binding to a 100‐bp promoter fragment of Slc7a5 ( n = 3). (H) Predicted Tcf1 binding site in the Slc7a5 promoter fragment identified using JASPAR. (I). Splenic CD4 + T cells stimulated with anti‐CD3/CD28 antibodies for 24 h were infected with lentivirus carrying murine pCDH‐Tcf1 T289E , pCDH‐Tcf1 T289A or pCDH‐Tcf1. CD4 + T cells were collected 48 h post‐infection, and RT‐qPCR was performed to detect the mRNA expression levels of Tcf7 and Slc7a5 ( n = 3). (J) Splenic CD4 + T cells stimulated with anti‐CD3/CD28 antibodies for 24 h were infected with pCDH‐H1‐EF1‐Puro scramble control or murine Tcf1 shRNA retroviruses. RT‐qPCR was performed to detect the mRNA expression levels of Tcf7 and Slc7a5 ( n = 3). ** p < 0.001, *** p < 0.0001, **** p < 0.00001.

Article Snippet: The primary antibodies for immunoblot were: Tcf1 (cat# 2203s, cst), Mapk13 (cat# A7496, ABclonal), Slc7a5 (cat# A2833, ABclonal), Gapdh (cat# A19056, ABclonal), Mat2a (cat# A19272, ABclonal), H3K27me3 (cat# 9733, cst), H3K4me3 (cat# 9751, cst), H3 (cat# 4499, cst).

Techniques: Isolation, RNA Sequencing, Quantitative RT-PCR, Western Blot, ChIP-sequencing, Expressing, Binding Assay, Luciferase, Infection, Control, shRNA

Slc7a5 deletion induces chronic graft tolerance. (A) Construction of the CAV model in Slc7a5 fl/fl and Slc7a5 −/− recipient mice. (B) Survival curves of transplanted hearts in Slc7a5 fl/fl and Slc7a5 −/− recipient mice ( n = 6). (C–F) Transplanted hearts from Slc7a5 fl/fl and Slc7a5 −/− recipient mice were harvested on day 30 post‐heart transplantation. Morphological comparisons (C), representative histological staining (D), CAV scores (E), and representative mIHC staining (F) of transplanted hearts ( n = 6). (G–I) Representative flow cytometric plots and proportions of CD4 + T cells and CD8 + T cells in transplanted hearts, peripheral blood, and spleens of Slc7a5 fl/fl and Slc7a5 −/− recipient mice ( n = 6). ns, not significant, ** p < 0.001, **** p < 0.00001.

Journal: Advanced Science

Article Title: Targeting the Mapk13‐Tcf1‐Slc7a5 Axis via One‐Carbon Metabolic Regulation to Prevent Chronic Allograft Vasculopathy

doi: 10.1002/advs.202520815

Figure Lengend Snippet: Slc7a5 deletion induces chronic graft tolerance. (A) Construction of the CAV model in Slc7a5 fl/fl and Slc7a5 −/− recipient mice. (B) Survival curves of transplanted hearts in Slc7a5 fl/fl and Slc7a5 −/− recipient mice ( n = 6). (C–F) Transplanted hearts from Slc7a5 fl/fl and Slc7a5 −/− recipient mice were harvested on day 30 post‐heart transplantation. Morphological comparisons (C), representative histological staining (D), CAV scores (E), and representative mIHC staining (F) of transplanted hearts ( n = 6). (G–I) Representative flow cytometric plots and proportions of CD4 + T cells and CD8 + T cells in transplanted hearts, peripheral blood, and spleens of Slc7a5 fl/fl and Slc7a5 −/− recipient mice ( n = 6). ns, not significant, ** p < 0.001, **** p < 0.00001.

Techniques: Transplantation Assay, Staining

Methionine restriction impairs the stem‐like properties of CD4 + T cells. (A,B) Effects of Mapk13 deletion and Slc7a5 deletion on amino acid consumption from the culture media in CD4 + T cells ( n = 3). (C) CD4 + T cells were isolated from the spleen of Slc7a5 fl/fl and Slc7a5 −/− mice and stimulated with anti‐CD3/CD28 antibodies for 72 h. Cells were subsequently collected for RNA‐seq. Gene Set Enrichment Analysis (GSEA) of amino acid metabolism‐related KEGG signaling pathways ( n = 3). (D‐K) CD4 + T cells were isolated from the spleen and stimulated with anti‐CD3/CD28 antibodies for 72 h. Then the cells were collected for Western blotting or flow cytometry. (D) Cell proliferation of CD4 + T cells from Slc7a5 fl/fl and Slc7a5 −/− mice ( n = 6). (E) Cell proliferation of CD4 + T cells cultured in standard medium or MR medium ( n = 5). (F) Mat2a protein levels of CD4 + T cells from Slc7a5 fl/fl and Slc7a5 −/− mice ( n = 3). (G) Mat2a protein levels of CD4 + T cells cultured in standard medium or MR medium ( n = 3). (H) Cell proliferation of CD4 + T cells cultured in the presence of FIDAS‐5 or DMSO ( n = 5). (I) Cell proliferation of CD4 + T cells cultured in the presence of DZ2002 or DMSO ( n = 5). (J) Tcf1 protein levels of CD4 + T cells cultured in standard medium or MR medium ( n = 3). (K) Protein levels of H3K4me3 in CD4 + T cells from Mapk13 fl/fl and Mapk13 −/− mice ( n = 3). (L) CUT&Tag‐seq tracks of H3K4me3 binding at Tcf7 loci. (M) Protein levels of H3K27me3 in CD4 + T cells from Mapk13 fl/fl and Mapk13 −/− mice ( n = 3). (N) CUT&Tag‐seq tracks of H3K27me3 binding at Tcf7 loci. ns, not significant, * p < 0.01, ** p < 0.001, *** p < 0.0001, **** p < 0.00001.

Journal: Advanced Science

Article Title: Targeting the Mapk13‐Tcf1‐Slc7a5 Axis via One‐Carbon Metabolic Regulation to Prevent Chronic Allograft Vasculopathy

doi: 10.1002/advs.202520815

Figure Lengend Snippet: Methionine restriction impairs the stem‐like properties of CD4 + T cells. (A,B) Effects of Mapk13 deletion and Slc7a5 deletion on amino acid consumption from the culture media in CD4 + T cells ( n = 3). (C) CD4 + T cells were isolated from the spleen of Slc7a5 fl/fl and Slc7a5 −/− mice and stimulated with anti‐CD3/CD28 antibodies for 72 h. Cells were subsequently collected for RNA‐seq. Gene Set Enrichment Analysis (GSEA) of amino acid metabolism‐related KEGG signaling pathways ( n = 3). (D‐K) CD4 + T cells were isolated from the spleen and stimulated with anti‐CD3/CD28 antibodies for 72 h. Then the cells were collected for Western blotting or flow cytometry. (D) Cell proliferation of CD4 + T cells from Slc7a5 fl/fl and Slc7a5 −/− mice ( n = 6). (E) Cell proliferation of CD4 + T cells cultured in standard medium or MR medium ( n = 5). (F) Mat2a protein levels of CD4 + T cells from Slc7a5 fl/fl and Slc7a5 −/− mice ( n = 3). (G) Mat2a protein levels of CD4 + T cells cultured in standard medium or MR medium ( n = 3). (H) Cell proliferation of CD4 + T cells cultured in the presence of FIDAS‐5 or DMSO ( n = 5). (I) Cell proliferation of CD4 + T cells cultured in the presence of DZ2002 or DMSO ( n = 5). (J) Tcf1 protein levels of CD4 + T cells cultured in standard medium or MR medium ( n = 3). (K) Protein levels of H3K4me3 in CD4 + T cells from Mapk13 fl/fl and Mapk13 −/− mice ( n = 3). (L) CUT&Tag‐seq tracks of H3K4me3 binding at Tcf7 loci. (M) Protein levels of H3K27me3 in CD4 + T cells from Mapk13 fl/fl and Mapk13 −/− mice ( n = 3). (N) CUT&Tag‐seq tracks of H3K27me3 binding at Tcf7 loci. ns, not significant, * p < 0.01, ** p < 0.001, *** p < 0.0001, **** p < 0.00001.

Techniques: Isolation, RNA Sequencing, Protein-Protein interactions, Western Blot, Flow Cytometry, Cell Culture, Binding Assay

(A) Schematic representation of SLC3A2-containing amino acid transporters. (B) MDA-MB-231 and (C) PANC1 cells were seeded on 2 mg/ml collagen I (Coll) or plastic (P) under complete or Glc starvation media for 24 hours, mRNA was extracted and the levels of SLC3A2, SLC7A5, SLC7A6, SLC7A8 and GAPDH as housekeeping gene were quantified by SYBR-green qPCR. Data are presented as 2 -ΔCT ; mean ± SEM, N=3 independent experiments, ****p < 0.0001 Kruskal-Wallis, Dunn’s multiple comparisons test. (D,E) MDA-MB-231 and (F,G) PANC1 cells were seeded on 2 mg/ml collagen I (Coll) or plastic (P) under Glc starvation for one or two days, respectively. Cells were fixed and stained with (D,F) Hoechst 33342 (magenta), SLC3A2 (cyan) and Phalloidin Alexa Fluor 555 (grey) or (E,G) Hoechst 33342 (cyan), SLC7A5 (red) and Phalloidin Alexa Fluor 555 (grey). Bar, 20 µm. Images were collected with a ZEISS LSM98O Airyscan2 microscope and analysed by Fiji/lmageJ software. Data are presented as mean ± SEM, N=3 independent experiments (the bigger dots represent mean intensity of each field of view); **p < 0.01, ***p < 0.001 Mann-Whitney test. (H, I) MDA-MB-231 and PANC1 cells were transfected with an siRNA targeting SLC3A2 (3A2), an siRNA targeting SLC7A5 (7A5) or a combination of siRNA targeting SLC3A2 and SLC7A5 (3A2+7A5) and grown on 2mg/ml collagen I under Glc starvation for one or three days, respectively. Metabolites were extracted and analysed by targeted mass spectrometry. N=3 independent experiments.

Journal: bioRxiv

Article Title: Collagen I promotes cancer cell survival via amino acid import and mTORC1 activation

doi: 10.1101/2025.11.26.690708

Figure Lengend Snippet: (A) Schematic representation of SLC3A2-containing amino acid transporters. (B) MDA-MB-231 and (C) PANC1 cells were seeded on 2 mg/ml collagen I (Coll) or plastic (P) under complete or Glc starvation media for 24 hours, mRNA was extracted and the levels of SLC3A2, SLC7A5, SLC7A6, SLC7A8 and GAPDH as housekeeping gene were quantified by SYBR-green qPCR. Data are presented as 2 -ΔCT ; mean ± SEM, N=3 independent experiments, ****p < 0.0001 Kruskal-Wallis, Dunn’s multiple comparisons test. (D,E) MDA-MB-231 and (F,G) PANC1 cells were seeded on 2 mg/ml collagen I (Coll) or plastic (P) under Glc starvation for one or two days, respectively. Cells were fixed and stained with (D,F) Hoechst 33342 (magenta), SLC3A2 (cyan) and Phalloidin Alexa Fluor 555 (grey) or (E,G) Hoechst 33342 (cyan), SLC7A5 (red) and Phalloidin Alexa Fluor 555 (grey). Bar, 20 µm. Images were collected with a ZEISS LSM98O Airyscan2 microscope and analysed by Fiji/lmageJ software. Data are presented as mean ± SEM, N=3 independent experiments (the bigger dots represent mean intensity of each field of view); **p < 0.01, ***p < 0.001 Mann-Whitney test. (H, I) MDA-MB-231 and PANC1 cells were transfected with an siRNA targeting SLC3A2 (3A2), an siRNA targeting SLC7A5 (7A5) or a combination of siRNA targeting SLC3A2 and SLC7A5 (3A2+7A5) and grown on 2mg/ml collagen I under Glc starvation for one or three days, respectively. Metabolites were extracted and analysed by targeted mass spectrometry. N=3 independent experiments.

Article Snippet: Primary antibodies for Phospho-S6 Ribosomal Protein ser235/236 and LC3A/B were from Cell Signalling; α2 integrin (CD49b)-FITC conjugated and β1 integrin (CD29)-Alexa Fluor 488 conjugated from BioLegend; α2 integrin for Western blotting from BD-bioscience; SLC3A2 (CD98) and SLC7A5 (LAT1) from Proteintech and GAPDH from SANTA CRUZ Biotechnology.

Techniques: SYBR Green Assay, Staining, Microscopy, Software, MANN-WHITNEY, Transfection, Mass Spectrometry

MDA-MB-231 (A-D) arid PANC1 (E-H) were transfected with an siRNA targeting SLC3A2 (3A2 si), an siRNA targeting SLC7A5 (7A5 si) or a non-targeting siRNA control (Nt si) and grown on 2 mg/ml collagen I under complete or Glc starvation media for 4 days. Cells were fixed and stained with Hoechst 33342. Images were collected by an ImageXpress micro and analysed by Meta×press software. Data are presented as mean ± SEM, N=3 independent experiments, *p < 0.05, **p < 0.01 and ****p < 0.0001 Mann-Whitney test. (I) MDA-MB-231-GFP cells were transfected with a combination of siRNAs targeting SLC3A2 and SLC7A5 (3A2+7A5 si) or a nontargeting siRNA control (Nt si) for 24 hours, spheroids were generated by the hanging drop method and embedded in 3 mg/ml collagen I and Geltrex (50:50 ratio) for 3 days under Glc starvation. Images were collected by a Nikon Confocal A1 microscope. Invasion area was quantified with Fiji/lmageJ. Bar, 200 µm. Data are presented as mean ± SEM, N=2 independent experiments. (J) PANC1 cells were transfected as in I, spheroids were generated by the hanging drop method and embedded in 3 mg/ml collagen land Geltrex (50:50 ratio) for 6 days under Glc starvation. Live images were collected every day by an Olympus E45O microscope. Bar, 250 µm. Data are presented as mean ± SEM, N=2 independent experiments. (K, L) E0771 mouse tumours organoids were grown in a 3 mg/ml collagen I and Geltrex (50:50) mixture and starved in Glc starvation media for 2 days in the presence or absence of 50 mM D-phenylalanine (D-Phe). Spheroids were imaged live every day by an Olympus E45O microscope. Bar, 250 µm. Data are presented as mean ± SEM, N=3 independent experiments, *p < 0.05, **p < 0.01 and ****p < 0.0001; (l,J,K) Two-way ANOVA, Tukey’s multiple comparisons test; (L) Mann-Whitney test.

Journal: bioRxiv

Article Title: Collagen I promotes cancer cell survival via amino acid import and mTORC1 activation

doi: 10.1101/2025.11.26.690708

Figure Lengend Snippet: MDA-MB-231 (A-D) arid PANC1 (E-H) were transfected with an siRNA targeting SLC3A2 (3A2 si), an siRNA targeting SLC7A5 (7A5 si) or a non-targeting siRNA control (Nt si) and grown on 2 mg/ml collagen I under complete or Glc starvation media for 4 days. Cells were fixed and stained with Hoechst 33342. Images were collected by an ImageXpress micro and analysed by Meta×press software. Data are presented as mean ± SEM, N=3 independent experiments, *p < 0.05, **p < 0.01 and ****p < 0.0001 Mann-Whitney test. (I) MDA-MB-231-GFP cells were transfected with a combination of siRNAs targeting SLC3A2 and SLC7A5 (3A2+7A5 si) or a nontargeting siRNA control (Nt si) for 24 hours, spheroids were generated by the hanging drop method and embedded in 3 mg/ml collagen I and Geltrex (50:50 ratio) for 3 days under Glc starvation. Images were collected by a Nikon Confocal A1 microscope. Invasion area was quantified with Fiji/lmageJ. Bar, 200 µm. Data are presented as mean ± SEM, N=2 independent experiments. (J) PANC1 cells were transfected as in I, spheroids were generated by the hanging drop method and embedded in 3 mg/ml collagen land Geltrex (50:50 ratio) for 6 days under Glc starvation. Live images were collected every day by an Olympus E45O microscope. Bar, 250 µm. Data are presented as mean ± SEM, N=2 independent experiments. (K, L) E0771 mouse tumours organoids were grown in a 3 mg/ml collagen I and Geltrex (50:50) mixture and starved in Glc starvation media for 2 days in the presence or absence of 50 mM D-phenylalanine (D-Phe). Spheroids were imaged live every day by an Olympus E45O microscope. Bar, 250 µm. Data are presented as mean ± SEM, N=3 independent experiments, *p < 0.05, **p < 0.01 and ****p < 0.0001; (l,J,K) Two-way ANOVA, Tukey’s multiple comparisons test; (L) Mann-Whitney test.

Techniques: Transfection, Control, Staining, Software, MANN-WHITNEY, Generated, Microscopy

(A,C) RNA sequencing data from basal-Like breast tumours (n=135) and normal breast tissue (n=291) or pancreatic adenocarcinoma tumours (n=179) and normal pancreatic tissue (n=171) for the co-expression of SLC3A2 and SLC7A5. *p< 0.05. Data were obtained from gepia2.com. (B) Overall survival of basal-like breast cancer patients with high (red) or low (blue) coexpression of SLC3A2 and SLC7A5. Data were obtained from gepia2.com. (D) Disease free survival of pancreatic adenocarcinoma patients with high (red) or low (blue) co-expression of SLC3A2 and SLC7A5. Data were obtained from gepia2.com. (E,F) RNA sequencing data and ROC analysis for the co-expression of SLC3A2 and SLC7A5 from chemoresistant (Non-responder) and chemosensitive (Responder) breast tumours. (E) p = 1,4e-6, Mann-Whitney test. (D) AUG = 0.628; ROC p value = 2.5e-07. Data were generated in ROCplot.com.

Journal: bioRxiv

Article Title: Collagen I promotes cancer cell survival via amino acid import and mTORC1 activation

doi: 10.1101/2025.11.26.690708

Figure Lengend Snippet: (A,C) RNA sequencing data from basal-Like breast tumours (n=135) and normal breast tissue (n=291) or pancreatic adenocarcinoma tumours (n=179) and normal pancreatic tissue (n=171) for the co-expression of SLC3A2 and SLC7A5. *p< 0.05. Data were obtained from gepia2.com. (B) Overall survival of basal-like breast cancer patients with high (red) or low (blue) coexpression of SLC3A2 and SLC7A5. Data were obtained from gepia2.com. (D) Disease free survival of pancreatic adenocarcinoma patients with high (red) or low (blue) co-expression of SLC3A2 and SLC7A5. Data were obtained from gepia2.com. (E,F) RNA sequencing data and ROC analysis for the co-expression of SLC3A2 and SLC7A5 from chemoresistant (Non-responder) and chemosensitive (Responder) breast tumours. (E) p = 1,4e-6, Mann-Whitney test. (D) AUG = 0.628; ROC p value = 2.5e-07. Data were generated in ROCplot.com.

Techniques: RNA Sequencing, Expressing, MANN-WHITNEY, Generated

Under Glc starvation, collagen I promoted α2β1 integrin-dependent mTORCI activation in breast and pancreatic cancer cells. Collagen I also increased the membrane levels of the LAT1-4F2hc amino acid transporter heavy chain, SLC3A2 (3A2), and light chain, SLC7A5 (7A5), stimulating amino acid import. This resulted in mTORCI activation and autophagy inhibition, leading to increased survival, proliferation and invasion, in 2D and 3C contexts.

Journal: bioRxiv

Article Title: Collagen I promotes cancer cell survival via amino acid import and mTORC1 activation

doi: 10.1101/2025.11.26.690708

Figure Lengend Snippet: Under Glc starvation, collagen I promoted α2β1 integrin-dependent mTORCI activation in breast and pancreatic cancer cells. Collagen I also increased the membrane levels of the LAT1-4F2hc amino acid transporter heavy chain, SLC3A2 (3A2), and light chain, SLC7A5 (7A5), stimulating amino acid import. This resulted in mTORCI activation and autophagy inhibition, leading to increased survival, proliferation and invasion, in 2D and 3C contexts.

Techniques: Activation Assay, Membrane, Inhibition

A Venn diagram illustrating the screening logic for SLC7A5. Integrated analysis of GSE153867 , GSE26712 , and OS databases identified nine genes highly expressed in drug-resistant cells with intersecting prognostic values. B Gene Expression Analysis. Analysis of the GSE153867 dataset revealed elevated SLC7A5 expression in Olaparib-resistant A2780 cells (C1–C8) compared to their parental controls (O1–O8). C WB. Western blot revealed higher SLC7A5 expression in HEY and SK-OV-3 cells compared to IOSE80, and lower expression in CAOV-3 and OVCAR-8 cells. D , E Drug Resistance Assay. IC50 values showed resistance to olaparib with high SLC7A5 expression. F WB. Western blot showed significant increases in SLC7A5 expression in olaparib-resistant cells. G Colony-formation assay. Knockdown of SLC7A5 significantly reduced Olaparib resistance in resistant cells. Left. HEY-R. Right. SK-OV-3-R. Plating 1000 cells per well with drug concentrations set at their respective IC50 values. H Statistical graph of Edu Assay. Edu incorporation assay showed that SLC7A5 knockdown significantly inhibited the proliferation of resistant cells treated with Olaparib. Edu incubation time was 2 h. Blue. DAPI, Green. Edu. Statistical graph displays Edu-positive rate in all cells. I Apoptosis Assay. SLC7A5 knockdown increased sensitivity to Olaparib in resistant cells. Cell apoptosis levels were detected using Annexin V and PI. J Colony-formation assay. The addition of leucine does not affect the regulation of olaparib resistance by SLC7A5. Plating 1000 cells per well with drug concentrations set at their respective IC50 values. Leucine (1 μM) treatment for 48 h. K Apoptosis Assay. SLC7A5’s regulation of olaparib resistance is leucine-independent. Cell apoptosis levels were detected using Annexin V and PI. L Statistical graph of Edu Assay. The addition of rapamycin does not affect the regulation of olaparib resistance by SLC7A5. Rapamycin (18.74 μM) treatment for 24 h. Edu incubation time was 2 h. Blue. DAPI, Green. Edu. Statistical graph displays Edu-positive rate in all cells. M Kaplan−Meier survival curves of nude mice implanted orthotopically with SK-OV-3 R cells. In a xenograft survival study conducted in nude mice, 5 million SK-OV-3 R cells were subcutaneously injected. Mice were divided into four groups, NC + vehivel, SLC7A5 + vehivel, NC + lue and SLC7A5 + leu. Once the tumors reached a volume of 50 mm³, each group was randomly divided into control and treatment subgroups. The treatment subgroup received olaparib every other day for a total of eight doses at 25 mg/kg. In accordance with animal ethics, mice were euthanized when tumor volumes reached 1000 mm³, which was recorded as the endpoint for survival. The median survival times were as follows. 40 days for the NC + vehivel group, 42.5 days for the NC + leu group, 20 days for the SLC7A5 + vehivel group, 17.5 days for the SLC7A5 + lue group (n = 10). N In vivo experiments. In vivo experiments demonstrated that SLC7A5 knockdown enhanced the inhibitory effect of Olaparib on ovarian cancer (n = 6). Upper panel. Schematic of in vivo treatment protocol. Lower panel. Tumor images. O Tumor weight (g). All in vitro findings are derived from a minimum of three independent experiments. Error bars denote the standard deviation. Statistical significance is represented as follows. * = P < 0.05, ** = P < 0.001, *** = P < 0.0001, ns = non-significant in comparison to normal or control treatment.

Journal: Oncogene

Article Title: SLC7A5-ERBB2 axis drives olaparib resistance via de novo lipid synthesis in ovarian cancer

doi: 10.1038/s41388-025-03584-w

Figure Lengend Snippet: A Venn diagram illustrating the screening logic for SLC7A5. Integrated analysis of GSE153867 , GSE26712 , and OS databases identified nine genes highly expressed in drug-resistant cells with intersecting prognostic values. B Gene Expression Analysis. Analysis of the GSE153867 dataset revealed elevated SLC7A5 expression in Olaparib-resistant A2780 cells (C1–C8) compared to their parental controls (O1–O8). C WB. Western blot revealed higher SLC7A5 expression in HEY and SK-OV-3 cells compared to IOSE80, and lower expression in CAOV-3 and OVCAR-8 cells. D , E Drug Resistance Assay. IC50 values showed resistance to olaparib with high SLC7A5 expression. F WB. Western blot showed significant increases in SLC7A5 expression in olaparib-resistant cells. G Colony-formation assay. Knockdown of SLC7A5 significantly reduced Olaparib resistance in resistant cells. Left. HEY-R. Right. SK-OV-3-R. Plating 1000 cells per well with drug concentrations set at their respective IC50 values. H Statistical graph of Edu Assay. Edu incorporation assay showed that SLC7A5 knockdown significantly inhibited the proliferation of resistant cells treated with Olaparib. Edu incubation time was 2 h. Blue. DAPI, Green. Edu. Statistical graph displays Edu-positive rate in all cells. I Apoptosis Assay. SLC7A5 knockdown increased sensitivity to Olaparib in resistant cells. Cell apoptosis levels were detected using Annexin V and PI. J Colony-formation assay. The addition of leucine does not affect the regulation of olaparib resistance by SLC7A5. Plating 1000 cells per well with drug concentrations set at their respective IC50 values. Leucine (1 μM) treatment for 48 h. K Apoptosis Assay. SLC7A5’s regulation of olaparib resistance is leucine-independent. Cell apoptosis levels were detected using Annexin V and PI. L Statistical graph of Edu Assay. The addition of rapamycin does not affect the regulation of olaparib resistance by SLC7A5. Rapamycin (18.74 μM) treatment for 24 h. Edu incubation time was 2 h. Blue. DAPI, Green. Edu. Statistical graph displays Edu-positive rate in all cells. M Kaplan−Meier survival curves of nude mice implanted orthotopically with SK-OV-3 R cells. In a xenograft survival study conducted in nude mice, 5 million SK-OV-3 R cells were subcutaneously injected. Mice were divided into four groups, NC + vehivel, SLC7A5 + vehivel, NC + lue and SLC7A5 + leu. Once the tumors reached a volume of 50 mm³, each group was randomly divided into control and treatment subgroups. The treatment subgroup received olaparib every other day for a total of eight doses at 25 mg/kg. In accordance with animal ethics, mice were euthanized when tumor volumes reached 1000 mm³, which was recorded as the endpoint for survival. The median survival times were as follows. 40 days for the NC + vehivel group, 42.5 days for the NC + leu group, 20 days for the SLC7A5 + vehivel group, 17.5 days for the SLC7A5 + lue group (n = 10). N In vivo experiments. In vivo experiments demonstrated that SLC7A5 knockdown enhanced the inhibitory effect of Olaparib on ovarian cancer (n = 6). Upper panel. Schematic of in vivo treatment protocol. Lower panel. Tumor images. O Tumor weight (g). All in vitro findings are derived from a minimum of three independent experiments. Error bars denote the standard deviation. Statistical significance is represented as follows. * = P < 0.05, ** = P < 0.001, *** = P < 0.0001, ns = non-significant in comparison to normal or control treatment.

Article Snippet: The antibodies used were ACLY ( 15421-1-AP, Proteintech, RRID: AB_2223741 ), SLC7A5( 28670-1-AP, Proteintech, RRID: AB_2918188 ), ERBB2( 84046-1-RR, Proteintech ), KLHL25 ( 67774-1-Ig, Proteintech, RRID: AB_2918539 ), CUL3( 83619-5-RR, Proteintech ), ubiquitin ( 10201-2-AP, Proteintech, RRID: AB_671515 ), GAPDH( 60004-1-Ig, Proteintech, RRID: AB_2107436 ), and Beta Actin ( 866009-1-Ig, Proteintech, RRID: AB_2687938 ).

Techniques: Gene Expression, Expressing, Western Blot, Colony Assay, Knockdown, EdU Assay, Incubation, Apoptosis Assay, Injection, Control, In Vivo, In Vitro, Derivative Assay, Standard Deviation, Comparison

A Immunohistochemistry (IHC). IHC revealed elevated SLC7A5 expression in late-stage patients, which was positively correlated with pathological grade. B Statistical graph of IHC. The correlation between SLC7A5 expression and tumor staging was analyzed in tissue microarray samples. Results showed that SLC7A5 expression increased with tumor progression. Stage I (3/8), Stage II (9/34), Stage III (36/64), and Stage IV (25/26). This suggests a potential role of SLC7A5 in advanced tumor development. C Survival Analysis. Kaplan-Meier analysis indicated that patients with high SLC7A5 expression had poorer prognoses and shorter survival times. D , E WB. The high expression efficiency of SLC7A5 and the knockdown efficiency using lentiviral shSLC7A5 #1 and #2 were detected in HEY-R and SK-OV-3-R cells. Overexpression was achieved using SLC7A5 plasmid in CAOV-3 and OVCAR-8 cells. F , G Colony-formation assay. SLC7A5 knockdown significantly inhibited cell proliferation, whereas overexpression promoted it. F HEY-R and SK-OV-3-R, G Caov-3 and Ovcar-8. Plating 2000 cells per well. H , I Apoptosis Assays. SLC7A5 knockdown increased apoptosis in cells, whereas overexpression SLC7A5 inhibited it. H HEY-R and SK-OV-3-R, I Caov-3 and Ovcar-8. J – L In vivo experiments. Subcutaneous tumor growth in nude mice with SK-OV-3-R cells showed that SLC7A5 knockdown inhibited tumor growth. J Bioluminescence Imaging. K Tumor volume (mm 3 ). M IHC Analysis. IHC showed a significant decrease in Ki67 expression following SLC7A5 knockdown (n = 6). All the in vitro data were obtained from at least three independent experiments. Error bars represent the standard deviation. Statistical significance is denoted as follows. * P < 0.05, ** P < 0.001, *** P < 0.0001, ns non-significant compared to the normal or control treatments.

Journal: Oncogene

Article Title: SLC7A5-ERBB2 axis drives olaparib resistance via de novo lipid synthesis in ovarian cancer

doi: 10.1038/s41388-025-03584-w

Figure Lengend Snippet: A Immunohistochemistry (IHC). IHC revealed elevated SLC7A5 expression in late-stage patients, which was positively correlated with pathological grade. B Statistical graph of IHC. The correlation between SLC7A5 expression and tumor staging was analyzed in tissue microarray samples. Results showed that SLC7A5 expression increased with tumor progression. Stage I (3/8), Stage II (9/34), Stage III (36/64), and Stage IV (25/26). This suggests a potential role of SLC7A5 in advanced tumor development. C Survival Analysis. Kaplan-Meier analysis indicated that patients with high SLC7A5 expression had poorer prognoses and shorter survival times. D , E WB. The high expression efficiency of SLC7A5 and the knockdown efficiency using lentiviral shSLC7A5 #1 and #2 were detected in HEY-R and SK-OV-3-R cells. Overexpression was achieved using SLC7A5 plasmid in CAOV-3 and OVCAR-8 cells. F , G Colony-formation assay. SLC7A5 knockdown significantly inhibited cell proliferation, whereas overexpression promoted it. F HEY-R and SK-OV-3-R, G Caov-3 and Ovcar-8. Plating 2000 cells per well. H , I Apoptosis Assays. SLC7A5 knockdown increased apoptosis in cells, whereas overexpression SLC7A5 inhibited it. H HEY-R and SK-OV-3-R, I Caov-3 and Ovcar-8. J – L In vivo experiments. Subcutaneous tumor growth in nude mice with SK-OV-3-R cells showed that SLC7A5 knockdown inhibited tumor growth. J Bioluminescence Imaging. K Tumor volume (mm 3 ). M IHC Analysis. IHC showed a significant decrease in Ki67 expression following SLC7A5 knockdown (n = 6). All the in vitro data were obtained from at least three independent experiments. Error bars represent the standard deviation. Statistical significance is denoted as follows. * P < 0.05, ** P < 0.001, *** P < 0.0001, ns non-significant compared to the normal or control treatments.

Techniques: Immunohistochemistry, Expressing, Microarray, Knockdown, Over Expression, Plasmid Preparation, Colony Assay, In Vivo, Imaging, In Vitro, Standard Deviation, Control

A GSEA . The enrichment analysis results of gene SLC7A5 in the fatty acyl-CoA biosynthesis pathway highlight its potential significance. With a normalized enrichment score (NES) of 1.5658, a P-value of 0.031, and a false discovery rate (FDR) of 0.0927, it suggests that SLC7A5 may play a crucial role in this biosynthetic process, warranting further investigation and study. B Correlation Analysis. Analysis of TCGA database revealed a positive correlation between SLC7A5 and de novo lipid metabolism pathway-associated proteins ACLY, ACACA, and FASN, suggesting that SLC7A5 is involved in lipid metabolism remodeling. C ELISA. Measurements of Tri-Glycerides, Free Fatty Acids, and Total Cholesterol showed that SLC7A5 knockdown significantly inhibited lipid metabolism in HEY and SK-OV-3 cells. Cells (5 × 10 6 , lysed by sonication. D Lipid Droplet Staining. LD540 staining revealed reduced lipid droplets upon SLC7A5 knockdown. Red fluorescence was observed using a fluorescence microscope at an excitation wavelength of 537 nm. E Immunohistochemistry (IHC). IHC analysis of tumor samples from mice showing reduced ACLY expression following SLC7A5 knockdown. F ELISA. ELISA analysis revealed that SB 204990, an ACLY inhibitor, significantly inhibited lipid metabolism remodeling caused by SLC7A5 overexpression in CAOV-3 and OVCAR-8 cells. Cells were treated with 100 µM SB 204990 for 24 h. G Lipid Droplet Staining. LD540 staining shows a significant reduction in lipid droplet accumulation induced by SLC7A5 overexpression following treatment with SB 204990. H Western Blot Analysis. SB 204990 inhibited the upregulation of ACLY, ACACA, and FASN proteins induced by SLC7A5 overexpression. I – K SB 204990 treatment markedly suppressed the increased Olaparib resistance in CAOV-3 and OVCAR-8 cells caused by SLC7A5 overexpression. I Colony formation assay. J Cell viability assay. K Apoptosis Assay. All in vitro data were derived from at least three independent experiments. Error bars represent standard deviation. Statistical significance is indicated by the following symbols. * P < 0.05, ** P < 0.001, *** P < 0.0001, ns non-significant compared to the normal or control treatments.

Journal: Oncogene

Article Title: SLC7A5-ERBB2 axis drives olaparib resistance via de novo lipid synthesis in ovarian cancer

doi: 10.1038/s41388-025-03584-w

Figure Lengend Snippet: A GSEA . The enrichment analysis results of gene SLC7A5 in the fatty acyl-CoA biosynthesis pathway highlight its potential significance. With a normalized enrichment score (NES) of 1.5658, a P-value of 0.031, and a false discovery rate (FDR) of 0.0927, it suggests that SLC7A5 may play a crucial role in this biosynthetic process, warranting further investigation and study. B Correlation Analysis. Analysis of TCGA database revealed a positive correlation between SLC7A5 and de novo lipid metabolism pathway-associated proteins ACLY, ACACA, and FASN, suggesting that SLC7A5 is involved in lipid metabolism remodeling. C ELISA. Measurements of Tri-Glycerides, Free Fatty Acids, and Total Cholesterol showed that SLC7A5 knockdown significantly inhibited lipid metabolism in HEY and SK-OV-3 cells. Cells (5 × 10 6 , lysed by sonication. D Lipid Droplet Staining. LD540 staining revealed reduced lipid droplets upon SLC7A5 knockdown. Red fluorescence was observed using a fluorescence microscope at an excitation wavelength of 537 nm. E Immunohistochemistry (IHC). IHC analysis of tumor samples from mice showing reduced ACLY expression following SLC7A5 knockdown. F ELISA. ELISA analysis revealed that SB 204990, an ACLY inhibitor, significantly inhibited lipid metabolism remodeling caused by SLC7A5 overexpression in CAOV-3 and OVCAR-8 cells. Cells were treated with 100 µM SB 204990 for 24 h. G Lipid Droplet Staining. LD540 staining shows a significant reduction in lipid droplet accumulation induced by SLC7A5 overexpression following treatment with SB 204990. H Western Blot Analysis. SB 204990 inhibited the upregulation of ACLY, ACACA, and FASN proteins induced by SLC7A5 overexpression. I – K SB 204990 treatment markedly suppressed the increased Olaparib resistance in CAOV-3 and OVCAR-8 cells caused by SLC7A5 overexpression. I Colony formation assay. J Cell viability assay. K Apoptosis Assay. All in vitro data were derived from at least three independent experiments. Error bars represent standard deviation. Statistical significance is indicated by the following symbols. * P < 0.05, ** P < 0.001, *** P < 0.0001, ns non-significant compared to the normal or control treatments.

Techniques: Enzyme-linked Immunosorbent Assay, Knockdown, Sonication, Staining, Fluorescence, Microscopy, Immunohistochemistry, Expressing, Over Expression, Western Blot, Colony Assay, Viability Assay, Apoptosis Assay, In Vitro, Derivative Assay, Standard Deviation, Control

$A GSEA . The analysis shows a significant enrichment with a high NES of 2.0577, a P-value less than 0.001, and an FDR of 0.0119, indicating a strong association between SLC7A5 and this signaling pathway. B Correlation Analysis. TCGA database analysis revealed a positive correlation between ERBB2 and de novo lipid metabolism pathway-associated proteins ACLY, ACACA, and FASN, suggesting ERBB2 is involvement in lipid metabolism remodeling. C Proteasome Inhibition Assay. Knockdown of ERBB2 significantly reduces ACLY levels (line 2), which are restored upon treatment with 50 µM MG-132 for 6 h (line 4). D CHX Half-life Assay. ERBB2 knockdown shortens the half-life of ACLY (Left). Nuclear-cytoplasmic fractionation revealed that ERBB2 signaling selectively regulates ACLY activity/modification in the cytoplasmic compartment (Right). CHX. 10 µg/mL. E Denaturing IP Assay. Overexpression of ERBB2 decreases ACLY ubiquitination. F Co-Immunoprecipitation (Co-IP) Assay. Overexpression of ERBB2 enhances binding between CUL3 and ERBB2, while reducing binding between CUL3 and KLHL25. All in vitro data are derived from at least three independent experiments. G Proteasome Inhibition Assay. SLC7A5 knockdown increases ACLY (line 2), but ERBB2 knockdown further decreases ACLY levels (line 3). This decrease is inhibited by 50 µM MG-132 for 6 h (line 6). H Denaturing IP Assay. ERBB2 knockdown increases ACLY ubiquitination (line 2), which is reduced upon additional knockdown of CUL3(line 3) or KLHL25 (line 4). I Proteasome Inhibition Assay. ERBB2 knockdown decreases ACLY (line 2), but this decrease is reversed by knockdown of CUL3(line 3) or KLHL25 (line 4). MG-132 treatment (50 µM for 6 h) restores ACLY expression (line 7, 8). J Denaturing IP Assay. SLC7A5 overexpression reduces ACLY ubiquitination (line 2), which is increased upon ERBB2 knockdown (line 3). K ELISA. ELISA measurements of Tri-Glycerides, Free Fatty Acids, and Total Cholesterol showed that ERBB2 knockdown reverses the lipid metabolism remodeling caused by SLC7A5 overexpression. L Lipid Droplet Staining. LD540 staining demonstrated a reduction in lipid droplet accumulation induced by SLC7A5 following ERBB2 knockdown. Error bars represent standard deviation. Statistical significance is denoted as follows. * = P < 0.05, ** = P < 0.001, *** = P < 0.0001, ns = non-significant compared to normal or control treatment.$

Journal: Oncogene

Article Title: SLC7A5-ERBB2 axis drives olaparib resistance via de novo lipid synthesis in ovarian cancer

doi: 10.1038/s41388-025-03584-w

Figure Lengend Snippet: A GSEA . The analysis shows a significant enrichment with a high NES of 2.0577, a P-value less than 0.001, and an FDR of 0.0119, indicating a strong association between SLC7A5 and this signaling pathway. B Correlation Analysis. TCGA database analysis revealed a positive correlation between ERBB2 and de novo lipid metabolism pathway-associated proteins ACLY, ACACA, and FASN, suggesting ERBB2 is involvement in lipid metabolism remodeling. C Proteasome Inhibition Assay. Knockdown of ERBB2 significantly reduces ACLY levels (line 2), which are restored upon treatment with 50 µM MG-132 for 6 h (line 4). D CHX Half-life Assay. ERBB2 knockdown shortens the half-life of ACLY (Left). Nuclear-cytoplasmic fractionation revealed that ERBB2 signaling selectively regulates ACLY activity/modification in the cytoplasmic compartment (Right). CHX. 10 µg/mL. E Denaturing IP Assay. Overexpression of ERBB2 decreases ACLY ubiquitination. F Co-Immunoprecipitation (Co-IP) Assay. Overexpression of ERBB2 enhances binding between CUL3 and ERBB2, while reducing binding between CUL3 and KLHL25. All in vitro data are derived from at least three independent experiments. G Proteasome Inhibition Assay. SLC7A5 knockdown increases ACLY (line 2), but ERBB2 knockdown further decreases ACLY levels (line 3). This decrease is inhibited by 50 µM MG-132 for 6 h (line 6). H Denaturing IP Assay. ERBB2 knockdown increases ACLY ubiquitination (line 2), which is reduced upon additional knockdown of CUL3(line 3) or KLHL25 (line 4). I Proteasome Inhibition Assay. ERBB2 knockdown decreases ACLY (line 2), but this decrease is reversed by knockdown of CUL3(line 3) or KLHL25 (line 4). MG-132 treatment (50 µM for 6 h) restores ACLY expression (line 7, 8). J Denaturing IP Assay. SLC7A5 overexpression reduces ACLY ubiquitination (line 2), which is increased upon ERBB2 knockdown (line 3). K ELISA. ELISA measurements of Tri-Glycerides, Free Fatty Acids, and Total Cholesterol showed that ERBB2 knockdown reverses the lipid metabolism remodeling caused by SLC7A5 overexpression. L Lipid Droplet Staining. LD540 staining demonstrated a reduction in lipid droplet accumulation induced by SLC7A5 following ERBB2 knockdown. Error bars represent standard deviation. Statistical significance is denoted as follows. * = P < 0.05, ** = P < 0.001, *** = P < 0.0001, ns = non-significant compared to normal or control treatment.

Techniques: Inhibition, Knockdown, Fractionation, Activity Assay, Modification, Over Expression, Ubiquitin Proteomics, Co-Immunoprecipitation Assay, Binding Assay, In Vitro, Derivative Assay, Expressing, Enzyme-linked Immunosorbent Assay, Staining, Standard Deviation, Control

A – C Cell Function Experiments . ERBB2 knockdown reverses olaparib resistance induced by SLC7A5 overexpression. A Clonogenic Assay. B Apoptosis Assay. C Cell Viability Assay. D , E Kaplan−Meier survival curves of nude mice implanted orthotopically with CAOV-3 cells. In a xenograft survival study conducted in nude mice, 5 million CAOV-3 cells were subcutaneously injected. D Mice were divided into three groups, NC + shCtrl, SLC7A5 + shCtrl, and shSLC7A5 + shERBB2, with 20 mice per group. Once the tumors reached a volume of 50 mm³, each group was randomly divided into control and treatment subgroups. The treatment subgroup received olaparib every other day for a total of eight doses at 25 mg/kg, while the control group received an equivalent volume of DMSO. In accordance with animal ethics, mice were euthanized when tumor volumes reached 1000 mm³, which was recorded as the endpoint for survival. The median survival times were as follows. 27.5 days for the (NC + shCtrl + DMSO) group, 15 days for the (SLC7A5 + shCtrl + DMSO) group, 30 days for the (SLC7A5 + shERBB2 + DMSO) group, 45 days for the (NC + shCtrl + Olaparib) group, 20 days for the (SLC7A5 + shCtrl + Olaparib) group, and 47.5 days for the (NC + shCtrl +Olaparib) group (n = 10). E The relative extension of survival was statistically analyzed following olaparib treatment. All the in vitro data were obtained from at least three independent experiments. Error bars represent standard deviation. * = P < 0.05, ** = P < 0.001, *** = P < 0.0001, ns = non-significant compared to normal or control treatment. F Clonogenic Assay . The knockdown of ERBB2 inhibits the proliferative effects induced by the overexpression of SLC7A5. G – K In vivo experiments. Subcutaneous implantation of CAOV-3 cells showed that ERBB2 inhibits SLC7A5-mediated tumor promotion. G Tumor. H Tumor Volume(mm 3 ). I Tumor Weight (g). J Immunohistochemistry (IHC). K Western Blot Analysis. All in vitro data points are derived from at least three independent experiments. Error bars represent the standard deviation. Statistical significance is indicated by the following symbols. * = P < 0.05, ** = P < 0.001, *** = P < 0.0001, ns = non-significant compared to normal or control treatment.

Journal: Oncogene

Article Title: SLC7A5-ERBB2 axis drives olaparib resistance via de novo lipid synthesis in ovarian cancer

doi: 10.1038/s41388-025-03584-w

Figure Lengend Snippet: A – C Cell Function Experiments . ERBB2 knockdown reverses olaparib resistance induced by SLC7A5 overexpression. A Clonogenic Assay. B Apoptosis Assay. C Cell Viability Assay. D , E Kaplan−Meier survival curves of nude mice implanted orthotopically with CAOV-3 cells. In a xenograft survival study conducted in nude mice, 5 million CAOV-3 cells were subcutaneously injected. D Mice were divided into three groups, NC + shCtrl, SLC7A5 + shCtrl, and shSLC7A5 + shERBB2, with 20 mice per group. Once the tumors reached a volume of 50 mm³, each group was randomly divided into control and treatment subgroups. The treatment subgroup received olaparib every other day for a total of eight doses at 25 mg/kg, while the control group received an equivalent volume of DMSO. In accordance with animal ethics, mice were euthanized when tumor volumes reached 1000 mm³, which was recorded as the endpoint for survival. The median survival times were as follows. 27.5 days for the (NC + shCtrl + DMSO) group, 15 days for the (SLC7A5 + shCtrl + DMSO) group, 30 days for the (SLC7A5 + shERBB2 + DMSO) group, 45 days for the (NC + shCtrl + Olaparib) group, 20 days for the (SLC7A5 + shCtrl + Olaparib) group, and 47.5 days for the (NC + shCtrl +Olaparib) group (n = 10). E The relative extension of survival was statistically analyzed following olaparib treatment. All the in vitro data were obtained from at least three independent experiments. Error bars represent standard deviation. * = P < 0.05, ** = P < 0.001, *** = P < 0.0001, ns = non-significant compared to normal or control treatment. F Clonogenic Assay . The knockdown of ERBB2 inhibits the proliferative effects induced by the overexpression of SLC7A5. G – K In vivo experiments. Subcutaneous implantation of CAOV-3 cells showed that ERBB2 inhibits SLC7A5-mediated tumor promotion. G Tumor. H Tumor Volume(mm 3 ). I Tumor Weight (g). J Immunohistochemistry (IHC). K Western Blot Analysis. All in vitro data points are derived from at least three independent experiments. Error bars represent the standard deviation. Statistical significance is indicated by the following symbols. * = P < 0.05, ** = P < 0.001, *** = P < 0.0001, ns = non-significant compared to normal or control treatment.

Techniques: Cell Function Assay, Knockdown, Over Expression, Clonogenic Assay, Apoptosis Assay, Viability Assay, Injection, Control, In Vitro, Standard Deviation, In Vivo, Immunohistochemistry, Western Blot, Derivative Assay

$A RT-PCR and WB Analysis. The knockout of SLC7A5 significantly inhibited the expression of ERBB2. Line left. HEY, Line right. SKOV3. B RT-PCR Analysis. RT-PCR showed that ELK1 knockdown significantly inhibits SLC7A5-mediated ERBB2 transcription. C Western Blot Analysis. Western blot demonstrated that ELK1 knockdown reverses the increase in ERBB2 protein levels caused by SLC7A5. D Nuclear and Cytoplasmic Fractionation. Nuclear and cytoplasmic fractionation combined with Western blot showed that SLC7A5 overexpression significantly promotes the nuclear translocation of pELK1. E Co-Immunoprecipitation (Co-IP). Co-IP confirmed SLC7A5- ELK1 protein-protein interactions. F Dual Luciferase Reporter Assay. This assay showed that SLC7A5 enhances ELK1 binding to the ERBB2 transcript in a dose-dependent manner. H RT-PCR Analysis. RT-PCR demonstrated that TDE significantly inhibits SLC7A5-mediated ERBB2 protein expression. G Dual Luciferase Reporter Assay. This assay showed that TDE significantly inhibits ELK1 binding to the ERBB2 transcript in a dose-dependent manner. I Chromatin Immunoprecipitation (ChIP) Assay. ChIP analysis showed that TDE significantly inhibits ELK1 binding to the ERBB2 transcript. J Dual Luciferase Reporter Assay. This assay demonstrated that ELK1 binds to the ERBB2 promoter sequence “CCTTCCATC”. Left. Schematic of dual luciferase reporter site mutations. Right. Dual luciferase assay results. K – M Proliferation Assay. TDE significantly mitigates the Olaparib resistance induced by SLC7A5 overexpression. K Edu Assay. L Colony-formation assay. M Cell Viability Test. All in vitro data are derived from at least three independent experiments. Error bars represent standard deviation. Statistical significance is denoted as follows. * = P < 0.05, ** = P < 0.001, *** = P < 0.0001, ns = non-significant compared to normal or control treatment.$

Journal: Oncogene

Article Title: SLC7A5-ERBB2 axis drives olaparib resistance via de novo lipid synthesis in ovarian cancer

doi: 10.1038/s41388-025-03584-w

Figure Lengend Snippet: A RT-PCR and WB Analysis. The knockout of SLC7A5 significantly inhibited the expression of ERBB2. Line left. HEY, Line right. SKOV3. B RT-PCR Analysis. RT-PCR showed that ELK1 knockdown significantly inhibits SLC7A5-mediated ERBB2 transcription. C Western Blot Analysis. Western blot demonstrated that ELK1 knockdown reverses the increase in ERBB2 protein levels caused by SLC7A5. D Nuclear and Cytoplasmic Fractionation. Nuclear and cytoplasmic fractionation combined with Western blot showed that SLC7A5 overexpression significantly promotes the nuclear translocation of pELK1. E Co-Immunoprecipitation (Co-IP). Co-IP confirmed SLC7A5- ELK1 protein-protein interactions. F Dual Luciferase Reporter Assay. This assay showed that SLC7A5 enhances ELK1 binding to the ERBB2 transcript in a dose-dependent manner. H RT-PCR Analysis. RT-PCR demonstrated that TDE significantly inhibits SLC7A5-mediated ERBB2 protein expression. G Dual Luciferase Reporter Assay. This assay showed that TDE significantly inhibits ELK1 binding to the ERBB2 transcript in a dose-dependent manner. I Chromatin Immunoprecipitation (ChIP) Assay. ChIP analysis showed that TDE significantly inhibits ELK1 binding to the ERBB2 transcript. J Dual Luciferase Reporter Assay. This assay demonstrated that ELK1 binds to the ERBB2 promoter sequence “CCTTCCATC”. Left. Schematic of dual luciferase reporter site mutations. Right. Dual luciferase assay results. K – M Proliferation Assay. TDE significantly mitigates the Olaparib resistance induced by SLC7A5 overexpression. K Edu Assay. L Colony-formation assay. M Cell Viability Test. All in vitro data are derived from at least three independent experiments. Error bars represent standard deviation. Statistical significance is denoted as follows. * = P < 0.05, ** = P < 0.001, *** = P < 0.0001, ns = non-significant compared to normal or control treatment.

Techniques: Reverse Transcription Polymerase Chain Reaction, Knock-Out, Expressing, Knockdown, Western Blot, Fractionation, Over Expression, Translocation Assay, Immunoprecipitation, Co-Immunoprecipitation Assay, Protein-Protein interactions, Luciferase, Reporter Assay, Binding Assay, Chromatin Immunoprecipitation, Sequencing, Proliferation Assay, EdU Assay, Colony Assay, In Vitro, Derivative Assay, Standard Deviation, Control

Journal: Oncogene

Article Title: SLC7A5-ERBB2 axis drives olaparib resistance via de novo lipid synthesis in ovarian cancer

doi: 10.1038/s41388-025-03584-w

Figure Lengend Snippet: SLC7A5-ERBB2 axis drives olaparib resistance via de novo lipid synthesis in ovarian cancer.

Techniques:

Review

anti slc7a5 (Proteintech)

Structured Review

Images

Related Articles

Immunohistochemistry:

Staining:

Microarray:

Western Blot:

Incubation:

Cell-Signaling:

Similar Products

Image Search Results