monoclonal mouse α  (Proteintech)

Journal: Discover Oncology

Article Title: SLC3A2 as a key anoikis−related gene for prognosis and tumor microenvironment remodeling in melanoma

doi: 10.1007/s12672-025-03125-7

Figure Lengend Snippet: Composition of genes within the anoikis-related prognostic model.The weight coefficients of genes included in the final model, with SLC3A2 displaying the strongest positive association with risk score

Article Snippet: To knock down SLC3A2 expression, specific short hairpin RNAs (shRNAs) targeting human SLC3A2 were designed and synthesized by GenePharma (Shanghai, China)(Supplemental Table 1).

Techniques:

Journal: Discover Oncology

Article Title: SLC3A2 as a key anoikis−related gene for prognosis and tumor microenvironment remodeling in melanoma

doi: 10.1007/s12672-025-03125-7

Figure Lengend Snippet: Expression, clinical significance, and biological implications of SLC3A2 in melanoma.( A–E ) SLC3A2 expression profiles across tumor and normal tissues, T stages, and treatment response groups. (F–J) Kaplan–Meier survival curves indicating the association between high SLC3A2 expression and worse prognosis in multiple datasets. ( K ) Forest plot of multivariate Cox regression analysis confirming SLC3A2 as an independent risk factor. ( L–O ) Gene set enrichment analysis (GSEA) identifying biological pathways significantly enriched in high SLC3A2 expression. ( P ) Drug sensitivity analysis highlighting potential compounds related to SLC3A2-mediated drug response

Article Snippet: To knock down SLC3A2 expression, specific short hairpin RNAs (shRNAs) targeting human SLC3A2 were designed and synthesized by GenePharma (Shanghai, China)(Supplemental Table 1).

Techniques: Expressing

Journal: Discover Oncology

Article Title: SLC3A2 as a key anoikis−related gene for prognosis and tumor microenvironment remodeling in melanoma

doi: 10.1007/s12672-025-03125-7

Figure Lengend Snippet: Functional validation of SLC3A2 in melanoma cell lines. ( A–B ) Wound healing and transwell migration assays showing impaired cell migration after SLC3A2 knockdown in A375 cells. ( C ) Colony formation assay demonstrating reduced proliferative capacity following SLC3A2 silencing

Article Snippet: To knock down SLC3A2 expression, specific short hairpin RNAs (shRNAs) targeting human SLC3A2 were designed and synthesized by GenePharma (Shanghai, China)(Supplemental Table 1).

Techniques: Functional Assay, Biomarker Discovery, Migration, Knockdown, Colony Assay

Journal: Journal of Cancer

Article Title: WTAP regulates Mitochondrial damage and Lipid oxidation in HCC by NOA1 mediated m6A modification.

doi: 10.7150/jca.102618

Figure Lengend Snippet: Figure 7. WTAP is independent of SLC7A11, not SLC3A2 or Xc- system in Huh-7 and SNU-449 cells. The siRNAs or plasmid were co-transfected to Huh-7 and SNU-449 cells by Lipofectamine™ 3000. 48h later, the total RNA or cell lysates were collected. RT-PCR and Western blot were used to detect the mRNA and protein levels of SLC7A11 (A-F) and SLC3A2 (G-L). *p < 0.05 compared with control groups.

Article Snippet: 16 https://www.jcancer.org 318 UK), anti-human SLC7A11 Rabbit mAb (#12691, Cell Signal Technology, USA), anti-human SLC3A2 Rabbit pAb (abs100601, Absin, China), anti-human NOA1 Rabbit pAb (103652-T32, Sino Biological, China), anti-human GSDMD Rabbit pAb (abs143289, Absin, China), Anti-human m6A antibody (No.202003, Synaptic Systems, Germany), anti-human β-actin Recombinant antibody (81115-1-RR, Proteintech, USA), anti-Rabbit IgG HRP-linked Antibody (#7074, Cell Signal Technology, USA), Peroxidase AffiniPure Goat Anti-Mouse IgG (H+L) (SA00001-1, Proteintech, USA).

Techniques: Plasmid Preparation, Transfection, Reverse Transcription Polymerase Chain Reaction, Western Blot, Control

Journal: Cell death and differentiation

Article Title: Anaplastic Lymphoma Kinase signaling stabilizes SLC3A2 expression via MARCH11 to promote neuroblastoma cell growth.

doi: 10.1038/s41418-024-01319-0

Figure Lengend Snippet: Fig. 3 Knockdown of SLC3A2 suppresses NB cell growth and increases retinoic acid-induced neuronal differentiation. A–D CLB-BAR, CLB- GE, and CLB-GAR cells were treated with either control siRNA or SLC3A2-targeting siRNAs. A Cell growth was monitored by Incucyte every 6 h for 72-120 h (n = 3, mean ± s.e.m.). B Lysates collected at 72 h after siRNA transfection were immunoblotted with anti-SLC3A2, pAKT(S473), AKT, pERK (T202/Y204), ERK, PARP (asterisk (*) marked cleaved PARP), γH2A.X, α-tubulin and GAPDH. GAPDH was employed as immunoblotting loading control. Quantification of cleaved PARP (PARP*) and γH2A.X were performed with ImageJ (n = 3), shown in Supplementary Fig. 3. C Cell cycle analysis performed by NucleoCounter NC-3000 after 72 h (n = 3, mean ± s.e.m.). D Representative images showing differentiation morphology of CLB-GE cells 72 h post siRNA transfection. Scale bar = 50 µm. Bar graph indicates percentage of differentiated cells following siRNA treatment (n = 5, mean ± s.e.m). E, F SK-N-BE(2) cells treated with either SLC3A2-targeting or control siRNAs for 24 h, followed by retinoic acid (RA, 5 µM) for 48 h. E Representative images showing differentiation morphology of SK-N-BE(2) cells. Scale bar = 50 µm. Bar graph indicates percentage of differentiated cells (n = 6, mean ± s.e.m.). F Immunoblotting for neuronal differentiation markers (RET, DLG2) in SK-N-BE(2) cells (n = 3). GAPDH was employed as immunoblotting loading control. Immunoblot experiments were performed in biologically independent triplicates. Quantification was performed with ImageJ and analyzed by Student t test (unpaired, two- tailed). p values are indicated (****P < 0.0001, ***0.0001 < P < 0.001, **0.001 < P < 0.01, *0.01 < P < 0.05, ns P ≥0.05); exact P-values are shown in Supplementary Fig. 3 and Supplementary Table 1.

Article Snippet: Primary antibodies against human SLC3A2 (#81977, was employed for immunoblotting, 1:2000; #13180, was used for immunoprecipitation), ALK (#3633, 1:2000), pALK (Y1278, #6941, 1:1000), ubiquitin (#3936, 1:1000), phospho-Tyrosine (p-Y-1000, #8954, 1:1000), pAKT (S473,#4060, 1:4000), AKT (#9272), pERK1/2 (Y204/T202, #4377, 1;2000), PARP (#9542, 1:1000), γH2A.X (Ser139, #9718, 1:1000), RET (#14556, 1:2000), DLG2 (#19046, 1:1000), SLC1A5 (ASCT2, #8057, 1:2000), SLC7A5 (LAT1, #32683, 1:1000), SLC7A11 (xCT, #12691, 1:1000), SLC38A1 (SNAT1, #36057, 1:2000), β-actin (#4970, 1:10000), GAPDH (#5174, 1:10,000) and α-tubulin (#2125, 1:10,000) were obtained from Cell Signaling Technology.

Techniques: Knockdown, Control, Transfection, Western Blot, Cell Cycle Assay, Two Tailed Test

Journal: Cell death and differentiation

Article Title: Anaplastic Lymphoma Kinase signaling stabilizes SLC3A2 expression via MARCH11 to promote neuroblastoma cell growth.

doi: 10.1038/s41418-024-01319-0

Figure Lengend Snippet: Fig. 5 Combined lorlatinib and AMXT-1501 treatment synergistically inhibits cell growth in ALK-driven NB and increases retinoic acid- induced neuronal differentiation. A IC50s of AMXT-1501 in CLB-BAR, CLB-GE, SK-N-BE(2) and SH-SY5Y cells after 5 d treatment (n = 3, mean ± s.e.m.). B CLB-BAR and CLB-GE cells were treated with lorlatinib and AMXT-1501 at the indicated concentrations and cell growth monitored by Incucyte every 6 h over a total of 120 h (n = 3, mean ± s.e.m.). The combination index value was calculated based on Chou- Talalay method using CompuSyn software [70]. C Immunoblot analysis of ALK signaling and apoptosis markers in CLB-GE cells treated with AMXT-1501 (0 or 8 µM) and lorlatinib (0 or 16 nM) as indicated for 24 h (n = 3). Lysates were immunoblotted with anti-ALK, pY1278-ALK, pAKT(S473), AKT, pERK (T202/Y204), ERK, PARP (asterisk (*) marked cleaved PARP), γH2A.X, α-tubulin and SLC3A2 and β-actin. β-actin was employed as immunoblotting loading control. Representative images of differentiation morphology of SK-N-BE(2) (D) and SH-SY5Y (E) cells after AMXT-1501 (0 or 8 µM) and RA (0 or 5 µM) treatment as indicated for 48 h or 24 h. Scale bar = 50 µm. Bar graph indicates percentage of differentiated cells (n = 3, mean ± s.e.m.). F Immunoblot analysis of neuronal differentiation markers (RET and DLG2) in SK-N-BE(2) and SH- SY5Y cells treated with AMXT-1501 (0 or 8 µM) and RA (0 or 5 µM) for 48 h or 24 h (n = 3). β-actin was employed as immunoblotting loading control. Immunoblot experiments (C and F) were performed in biologically independent triplicates. Quantifications were performed with ImageJ and analyzed by Student t-test (unpaired, two-tailed). p values are indicated (****P < 0.0001, ***0.0001 < P < 0.001, **0.001 < P < 0.01, *0.01 < P < 0.05, ns P ≥0.05); exact P values are shown in Supplementary Fig. 4 and Supplementary Table 1.

Article Snippet: Primary antibodies against human SLC3A2 (#81977, was employed for immunoblotting, 1:2000; #13180, was used for immunoprecipitation), ALK (#3633, 1:2000), pALK (Y1278, #6941, 1:1000), ubiquitin (#3936, 1:1000), phospho-Tyrosine (p-Y-1000, #8954, 1:1000), pAKT (S473,#4060, 1:4000), AKT (#9272), pERK1/2 (Y204/T202, #4377, 1;2000), PARP (#9542, 1:1000), γH2A.X (Ser139, #9718, 1:1000), RET (#14556, 1:2000), DLG2 (#19046, 1:1000), SLC1A5 (ASCT2, #8057, 1:2000), SLC7A5 (LAT1, #32683, 1:1000), SLC7A11 (xCT, #12691, 1:1000), SLC38A1 (SNAT1, #36057, 1:2000), β-actin (#4970, 1:10000), GAPDH (#5174, 1:10,000) and α-tubulin (#2125, 1:10,000) were obtained from Cell Signaling Technology.

Techniques: Software, Western Blot, Control, Two Tailed Test

Journal: Cell death and differentiation

Article Title: Anaplastic Lymphoma Kinase signaling stabilizes SLC3A2 expression via MARCH11 to promote neuroblastoma cell growth.

doi: 10.1038/s41418-024-01319-0

Figure Lengend Snippet: Fig. 6 ALK signaling upregulates amino acid transporter systems. Gene oncology GSEA analysis of RNA-seq datasets. A NB tumors from either Alk-F1178S;Th-MYCN mice or Rosa26_Alkal2;Th-MYCN mice compared to Th-MYCN mice. B NB1 cells treated with either ALKAL2 or/and lorlatinib for 24 h. C CLB-BAR cells treated with lorlatinib for 24 h compared to DMSO treated control. D Heat map indicating biotinylation of SLC1A5, SLC3A2, SLC6A15, SLC38A1 in SK-N-AS.ALK-BirA* expressing cells compared to BirA* controls. Columns represent three biological replicates, data from [41]. Immunoblotting for anti-pALK (Y1278), SLC1A5, SLC3A2, SLC7A5, SLC7A11, SLC38A1 and GAPDH in NB1 cells treated with ALKAL2 (1 µg/ml) and lorlatinib (0 or 30 nM) for 24 h (E) and CLB-BAR cells treated with lorlatinib (0, 30 nM) for 24 h (F). G Schematic visualization of the regulation of SLC3A2 downstream of ALK in NB cells (created with BioRender.com). Immunoblot experiments were performed in biologically independent triplicates. Quantifications were performed with ImageJ and analyzed by Student t test (unpaired, two- tailed). p values are indicated (****P < 0.0001, ***0.0001 < P < 0.001, **0.001 < P < 0.01, *0.01 < P < 0.05, ns P ≥0.05); exact P values are shown in Supplementary Fig. 5 and Supplementary Table 1.

Article Snippet: Primary antibodies against human SLC3A2 (#81977, was employed for immunoblotting, 1:2000; #13180, was used for immunoprecipitation), ALK (#3633, 1:2000), pALK (Y1278, #6941, 1:1000), ubiquitin (#3936, 1:1000), phospho-Tyrosine (p-Y-1000, #8954, 1:1000), pAKT (S473,#4060, 1:4000), AKT (#9272), pERK1/2 (Y204/T202, #4377, 1;2000), PARP (#9542, 1:1000), γH2A.X (Ser139, #9718, 1:1000), RET (#14556, 1:2000), DLG2 (#19046, 1:1000), SLC1A5 (ASCT2, #8057, 1:2000), SLC7A5 (LAT1, #32683, 1:1000), SLC7A11 (xCT, #12691, 1:1000), SLC38A1 (SNAT1, #36057, 1:2000), β-actin (#4970, 1:10000), GAPDH (#5174, 1:10,000) and α-tubulin (#2125, 1:10,000) were obtained from Cell Signaling Technology.

Techniques: RNA Sequencing, Control, Expressing, Western Blot, Two Tailed Test