antibody against slc3a2  (Proteintech)

Journal: Cancer Science

Article Title: Epigenetically‐Upregulated CD98 Shed Light on the Precancerous Diagnosis and Prognosis Prediction of Esophageal Cancer

doi: 10.1111/cas.70128

Figure Lengend Snippet: CD98 specific CpG unit methylation is negatively correlated with its expression and the prognosis of ESCC. (A) Comparison of CpG units methylation of CD98 promoter region in ESCC and normal samples. Each row represents one sample. Each column shows clusters of CpG units, which are individual CpG sites or combinations of CpG sites. The color gradient between red and blue indicates the methylation of each CD98 CpG unit in each sample, ranging from 0 to 1. Gray represents insufficient or missing data. (B) Methylation levels of 14 informative CpG sites in the CD98 promoter in normal and ESCC samples. * p < 0.05 (Mann–Whitney U test). (C) Analysis of scatterplots and simple linear regression graphically displaying the correlation between the methylation level of each CpG_23.24.25.26.27 and CD98 expression in ESCC samples by Spearman correlation coefficient analysis. (D) Kaplan–Meier survival curves of ESCC patients stratified by CD98 CpG_23.24.25.26.27 methylation.

Article Snippet: The primary antibody against CD98 was purchased from Proteintech (Chicago, USA) at a dilution of 1:1000, and the P53 and Ki67 antibodies were purchased from ZSGB‐BIO (Beijing, China) at a dilution of 1:50.

Techniques: Methylation, Expressing, Comparison, MANN-WHITNEY

Journal: Cancer Science

Article Title: Epigenetically‐Upregulated CD98 Shed Light on the Precancerous Diagnosis and Prognosis Prediction of Esophageal Cancer

doi: 10.1111/cas.70128

Figure Lengend Snippet: CD98 is highly expressed in ESCC and correlates with poor prognosis. (A) CD98 mRNA levels in different cancer types from TCGA data. (B) IHC analysis of the CD98 protein in adjacent normal and ESCC tissues. Right image magnification ×40; left image magnification ×200. (C) Quantification of positive staining for the CD98 protein in (B). (D, E) Distribution ratios of adjacent normal tissues and ESCC tissues in four‐level score (0–1, 2–3, 4–8 and 9–12) of CD98 expression. (F) Kaplan–Meier survival curves of ESCC patients stratified by CD98 expression. ACC, adrenocortical carcinoma; BLCA, bladder urothelial carcinoma; BRCA, breast invasive carcinoma; CESC, cervical squamous cell carcinoma and endocervical adeno carcinoma; CHOL, cholangio carcinoma; COAD, colon adenocarcinoma; DLBC, lymphoid neoplasm diffuse large B‐cell lymphoma; ESCA, esophageal carcinoma; GBM, glioblastoma multiforme; HNSC, head and neck squamous cell carcinoma; KICH, kidney chromophobe; KIRC, kidney renal clear cell carcinoma; KIRP, kidney renal papillary cell carcinoma; LAML, acute myeloid leukemia; LGG, brain lower grade glioma; LIHC, liver hepatocellular carcinoma; LUAD, lung adenocarcinoma; LUSC, lung squamous cell carcinoma; MESO, mesothelioma; OV, ovarian serous cystadenocarcinoma; PAAD, pancreatic adenocarcinoma; PCPG, pheochromocytoma and paraganglioma; PRAD, prostate adenocarcinoma; READ, rectum adenocarcinoma; SARC, sarcoma; SKCM, skin cutaneous melanoma; STAD, stomach adenocarcinoma; TGCT, testicular germ cell tumors; THCA, thyroid carcinoma; THYM, thymoma; UCEC, uterine corpus endometrial carcinoma; UCS, uterine carcinosarcoma; UVM, uveal melanoma.

Article Snippet: The primary antibody against CD98 was purchased from Proteintech (Chicago, USA) at a dilution of 1:1000, and the P53 and Ki67 antibodies were purchased from ZSGB‐BIO (Beijing, China) at a dilution of 1:50.

Techniques: Staining, Expressing

Journal: Cancer Science

Article Title: Epigenetically‐Upregulated CD98 Shed Light on the Precancerous Diagnosis and Prognosis Prediction of Esophageal Cancer

doi: 10.1111/cas.70128

Figure Lengend Snippet: Differential expression analysis of CD98 KD and control ESCC cell lines. (A) Western blotting showing the expression of CD98 in EC9706 and Eca109 cells transfected with RNAi negative control (control) or si‐CD98 (siRNA). (B) Heat map of TOP40 differentially expressed genes. (C) Circle diagram of the GO term analysis. The first circle indicates the top 20 GO terms; the second circle indicates the number of this GO term and the Q value in the background of differential proteins; and the third circle represents the bar chart of the proportion of upregulated and downregulated proteins. (D) Classification statistical bubble diagram of the KEGG enrichment (top 20). Different colors indicate different functional categories.

Article Snippet: The primary antibody against CD98 was purchased from Proteintech (Chicago, USA) at a dilution of 1:1000, and the P53 and Ki67 antibodies were purchased from ZSGB‐BIO (Beijing, China) at a dilution of 1:50.

Techniques: Quantitative Proteomics, Control, Western Blot, Expressing, Transfection, Negative Control, Functional Assay

Journal: Cancer Science

Article Title: Epigenetically‐Upregulated CD98 Shed Light on the Precancerous Diagnosis and Prognosis Prediction of Esophageal Cancer

doi: 10.1111/cas.70128

Figure Lengend Snippet: Knockdown of CD98 inhibits ESCC cell proliferation via G1 arrest. (A, B) Knockdown of CD98 inhibits cell growth as determined by colony formation assays. (C, D) CCK‐8 assays showed that silencing of endogenous CD98 inhibited cell proliferation. (E, F) Knockdown of CD98 induced cell cycle arrest at the G1 phase. * p < 0.05, ** p < 0.01.

Article Snippet: The primary antibody against CD98 was purchased from Proteintech (Chicago, USA) at a dilution of 1:1000, and the P53 and Ki67 antibodies were purchased from ZSGB‐BIO (Beijing, China) at a dilution of 1:50.

Techniques: Knockdown, CCK-8 Assay

Journal: Cancer Science

Article Title: Epigenetically‐Upregulated CD98 Shed Light on the Precancerous Diagnosis and Prognosis Prediction of Esophageal Cancer

doi: 10.1111/cas.70128

Figure Lengend Snippet: Correlation of CD98 distribution and expression patterns with ESCC progression. (A) Representative HE staining (left panel) and CD98 IHC staining (middle panel) in normal, LGIN, HGIN, and invasive carcinoma tissues. Scale bar, 50 μm. Cartoons demonstrating CD98 expression (CD98‐positive cells appear dark brown cell membranes) in the development of esophageal invasive carcinoma (right panel). Relationship between pathological changes and (B) CD98 and (C) P53 score. (D) SPSS correlation analysis between CD98 score and P53 score.

Article Snippet: The primary antibody against CD98 was purchased from Proteintech (Chicago, USA) at a dilution of 1:1000, and the P53 and Ki67 antibodies were purchased from ZSGB‐BIO (Beijing, China) at a dilution of 1:50.

Techniques: Expressing, Staining, Immunohistochemistry

Journal: Cancer Science

Article Title: Epigenetically‐Upregulated CD98 Shed Light on the Precancerous Diagnosis and Prognosis Prediction of Esophageal Cancer

doi: 10.1111/cas.70128

Figure Lengend Snippet: Expression patterns of CD98 and P53 in esophageal intraepithelial neoplasia. (A) Representative images showing four different expression patterns in LGIN tissues: CD98 + /P53 + (pattern 1), CD98 − /P53 + (pattern 2), CD98 + /P53 − (pattern 3), and CD98 − /P53 − (pattern 4). (B) Representative images showing four different expression patterns in HGIN tissues: CD98 + /P53 + (pattern 1), CD98 − /P53 + (pattern 2), CD98 + /P53 − (pattern 3), and CD98 − /P53 − (pattern 4). Scale bar, 50 μm. TOP panel: HE staining; middle panel: CD98 IHC staining; bottom panel: P53 IHC staining.

Article Snippet: The primary antibody against CD98 was purchased from Proteintech (Chicago, USA) at a dilution of 1:1000, and the P53 and Ki67 antibodies were purchased from ZSGB‐BIO (Beijing, China) at a dilution of 1:50.

Techniques: Expressing, Staining, Immunohistochemistry

Journal: Cancer Science

Article Title: Epigenetically‐Upregulated CD98 Shed Light on the Precancerous Diagnosis and Prognosis Prediction of Esophageal Cancer

doi: 10.1111/cas.70128

Figure Lengend Snippet: Schematic graph demonstrating different expression patterns of CD98 and P53 in the development of invasive esophageal carcinoma. CD98‐positive cells appeared as dark brown cell membranes and P53‐positive cells showed dark brown nuclei. In the occurrence and progression of esophageal squamous cell carcinoma, CD98 and P53 showed distinct combinatorial expression patterns, including CD98 + /P53 + (A), CD98 + /P53 − (B), and CD98 − /P53 + (C).

Article Snippet: The primary antibody against CD98 was purchased from Proteintech (Chicago, USA) at a dilution of 1:1000, and the P53 and Ki67 antibodies were purchased from ZSGB‐BIO (Beijing, China) at a dilution of 1:50.

Techniques: Expressing

Journal: International Journal of Oral Science

Article Title: Host-microbe computational proteomic landscape in oral cancer revealed key functional and metabolic pathways between Fusobacterium nucleatum and cancer progression

doi: 10.1038/s41368-024-00326-8

Figure Lengend Snippet: The L-Glutamate degradation pathway shows an association with bacterial peptides derived from F. nucleatum . a Scatter plots of quantitative value of SLC3A2 from the human proteomic dataset in Fig. . b Confocal image of the cystine/glutamate antiporter (System x c - ) showing the expression and colocalization of SLC3A2 (green) and SLC7A11 (red) in HSC3 cells. c Scatter plots of extracellular L-glutamate from HSC3 cell cultures in the presence or absence of the Imidazole Ketone Erastin (IKE) and d in HSC3 cells uninfected or infected with F. nucleatum for 24 h. e Western blot of SLC7A11 in the HSC3 cells uninfected or infected with F. nucleatum for 24 h. f Scatter plots and representative images or dot plots of bacterial infection in the absence or presence of IKE by Incucyte and g Flow cytometry. h Schematic overview of F. nucleatum influencing the System xc- and L-Glutamate efflux. For all statistical analysis T test was used, **** P < 0.000 1, *** P < 0.001, ** P < 0.01 and * P < 0.05 were considered significant

Article Snippet: 6 × 10 4 HSC3 cells were seeded onto glass coverslips for 24 h. Subsequently, the cells were fixed with 4% PFA for 30 min. Next, three washes were performed with 1X PBS, followed by overnight incubation with primary antibodies against SLC3A2 (Santa Cruz #SC-59145) and SLC7A11 (Cell Signaling #12691).

Techniques: Derivative Assay, Expressing, Infection, Western Blot, Flow Cytometry

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

Journal: Bioengineering & Translational Medicine

Article Title: Synergistic ferroptosis‐starvation therapy for bladder cancer based on hyaluronic acid modified metal–organic frameworks

doi: 10.1002/btm2.10515

Figure Lengend Snippet: ZPG@H caused cell death via ferroptosis. (a) Cell viabilities of T24 cells treated with ZPG@H (30 μg/mL) and different inhibitors (cell viabilities of cells only treated with PBS was set as 100%). (b) Fluorescence imaging of JC‐1 stained T24 cells treated with ZPG@H (30 μg/mL) and Fer‐1 (20 μM) for 6 h. The scale bar represents 200 μm. (c) Flow cytometry analysis of reactive oxygen species (ROS)‐probe‐stained T24 cells incubated with ZPG@H (30 μg/mL) and Fer‐1 (20 μM) for 6 h. (d) Fluorescence imaging of ROS‐probe‐stained T24 cells incubated with ZPG@H (30 μg/mL) and Fer‐1 (20 μM) for 6 h. (e) Flow cytometry analysis of JC‐1 stained T24 cells treated with ZPG@H (30 μg/mL) and Fer‐1 (20 μM) for 6 h. (f) Average PE‐Texas Red fluorescence intensities of ROS‐probe‐stained T24 cells incubated with ZPG@H (30 μg/mL) and Fer‐1 (20 μM) for 6 h. Asterisks indicate significant differences (* p < 0. 05; ** p < 0.01; *** p < 0.001). (g) Protein expression levels of SLC7A11, GPX4, and SLC3A2 after treated with different concentrations of ZPG@H for 24 h, 30 μg/mL ZPG@H for different time and with or without Fer‐1 (20 μM). GAPDH was used as an internal control.

Article Snippet: Then, membranes were incubated with antibodies against SLC7A11, SLC3A2, GPX4, and GAPDH (1:1000, Proteintech, China), followed by incubation with secondary antibodies.

Techniques: Fluorescence, Imaging, Staining, Flow Cytometry, Incubation, Expressing, Control