Journal: Cell Reports Medicine

Article Title: Synergistic T cell signaling by 41BB and CD28 is optimally achieved by membrane proximal positioning within parallel chimeric antigen receptors

doi: 10.1016/j.xcrm.2021.100457

Figure Lengend Snippet:

Article Snippet: Anti-human CD98 - PE-Vio770 , Miltenyi Biotec , Cat# 130-105-710, RRID: AB_2659686.

Techniques: Control, Recombinant, Staining, Transfection, Enzyme-linked Immunosorbent Assay, Luciferase, Gene Expression, Cloning, Software

Journal: Cells

Article Title: Echinococcus granulosus -Induced Liver Damage Through Ferroptosis in Rat Model

doi: 10.3390/cells14050328

Figure Lengend Snippet: (A) Ultra-structure of hepatocytes near cysts at different stages of infection with PSCs (0, 1, 3, and 6 months); mitochondria are indicated by white arrows,PSC are indicated by black arrows( n = 3 rats/group, scale bars represent 2 μm and 1 μm). ( B ) Fe 2+ , ( C ) GSH, ( D ( a , b )) ROS, ( E ) MDA, ( F ) SOD, and ( G ) LDH of hepatocytes near cysts at different stages of infection with PSCs. ( H ( a , b )) Western blotting of protein expression levels of TFRC, GPX4, FTH1, NOX1, SLC3A2, and SLC7A11 in ferroptosis signaling pathway in rat liver tissue at different stages of infection with PSCs ( n = 3 rats/group)); * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 (Student t test).

Article Snippet: The antibodies used in this study were as follows: TFRC (Boster, 1: 500, Rabbit, BA0462-2, Wuhan, China); Caspase-3 (Boster, 1:500, Rabbit, BA3257, Wuhan, China); GSDMD (Proteintech Group, 1:800, Rabbit, 20770-1-AP, Wuhan, China); LC3I/II (Abcam, 1:1000, Rabbit, ab192890, Shanghai, China); Anti-GPX4 Antibody (Boster, 1:400, Rabbit, BM5231,Wuhan, China); Anti-FTH1 Antibody (Boster, China, 1:400, Rabbit, BM4487,Wuhan, China); Anti-NOX1 Antibody (Boster, China, 1:400, Rabbit, BA3720,Wuhan, China); Anti-CD98 Antibody (SLC3A2, Boster, China, 1:400, Rabbit, A01794-1,Wuhan,China); Anti-xCT Antibody (SLC7A11,Abcam,1:400, Rabbit, A01794-1, Shanghai, China); β-actin (Sino Biological, 1:1000, Mouse, 100166-MM10, Beijing, China).

Techniques: Infection, Western Blot, Expressing

Journal: Cells

Article Title: Echinococcus granulosus -Induced Liver Damage Through Ferroptosis in Rat Model

doi: 10.3390/cells14050328

Figure Lengend Snippet: ( A ) Fe 2+ in normal BRL cells, PSCs and BRL co-cultured cells, and PSCs +Ferrostatin-1 and BRL co-cultured cells. ( B ) GSH, ( C ) GSSH, ( D ) GSH/GSSH, ( E ) SOD, ( F ) LDH, and ( G ) MDA concentration detection ( n = 6 rats/group). ( H ( a , b )) Western-blotting detection of expression levels of cell ferroptosis signaling pathway proteins in each group, such as TFRC, GPX4, FTH1, NOX1, SLC3A2, andSLC7A11 ( n = 3 rats/group); * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 (Student t test).

Article Snippet: The antibodies used in this study were as follows: TFRC (Boster, 1: 500, Rabbit, BA0462-2, Wuhan, China); Caspase-3 (Boster, 1:500, Rabbit, BA3257, Wuhan, China); GSDMD (Proteintech Group, 1:800, Rabbit, 20770-1-AP, Wuhan, China); LC3I/II (Abcam, 1:1000, Rabbit, ab192890, Shanghai, China); Anti-GPX4 Antibody (Boster, 1:400, Rabbit, BM5231,Wuhan, China); Anti-FTH1 Antibody (Boster, China, 1:400, Rabbit, BM4487,Wuhan, China); Anti-NOX1 Antibody (Boster, China, 1:400, Rabbit, BA3720,Wuhan, China); Anti-CD98 Antibody (SLC3A2, Boster, China, 1:400, Rabbit, A01794-1,Wuhan,China); Anti-xCT Antibody (SLC7A11,Abcam,1:400, Rabbit, A01794-1, Shanghai, China); β-actin (Sino Biological, 1:1000, Mouse, 100166-MM10, Beijing, China).

Techniques: Cell Culture, Concentration Assay, Western Blot, Expressing

Journal: Frontiers in Microbiology

Article Title: The CD147 Protein Complex Is Involved in Entry of Chikungunya Virus and Related Alphaviruses in Human Cells

doi: 10.3389/fmicb.2021.615165

Figure Lengend Snippet: CHIKV envelope association with the CD147 complex (A) Scheme of the used AP-MS approach. (B) CHIKV envelope Interaction partners of interest are listed. Examination of their plasma membrane presence (PM), presence of a transmembrane domain (TM), and MiST score (MiST) are indicated. Components of the CD147 protein complex retrieved in the CHIKV envelope affinity purification are indicated. (C–F) Western Blot of affinity purification of Strep-tagged Envelope protein. Detection of E2 and the interactors CD147, CD98, and SLC1A5. Affinity purification of untransfected cells (empty) were included as controls. On each blot input lysates of untransfected cells (empty) or E2 expressing cells (E2) and affinity purifications (AP) of the empty or E2 cells were loaded. For the CD147, CD98 and SLC1A5 blots 0.4 μl of input lysates and 10 μl of APs was loaded. For the E2 blot 5 μl of input lysates and 10 μl of APs was loaded.

Article Snippet: Membranes were washed with PBS-T and incubated with primary antibody anti-CD147 (ab232967, Abcam), anti-ASCT2 (SLC1A5) (V501, Cell Signaling), anti-CD98 (12206-T62, sino biologicals), anti-E1, anti-Strep (ab184224 or ab180957, Abcam) and anti-E2 (NR44002, Bei Resources) overnight at 4°C.

Techniques: Membrane, Affinity Purification, Western Blot, Expressing

Journal: Materials Today Bio

Article Title: A bioengineered tumor matrix-based scaffold for the evaluation of melatonin efficacy on head and neck squamous cancer stem cells

doi: 10.1016/j.mtbio.2024.101246

Figure Lengend Snippet: HNSCC microenvironment model. (A) Characterization of Cal-27 CSCs. Image of tumorspheres grown in suspension with serum-free spheres medium (10×). Cytometry histograms of CD98 and CD44 CSCs cell-membrane markers expression. Percentage of ALDH1 activity of cells grown in monolayer or in suspension (cytometry histograms besides). (B) Schematic representation of the TME components embedded in the hydrogel. Confocal representative images of cell viability of tumorspheres, FBs and MSCs (stroma) and the co-culture of stroma and tumorspheres (TME) cultured in the hydrogel. Living cells shown in green and nuclei of death cells in red. Scale bar: 200 μm. (C) Proliferation assay of the tumorspheres, FBs and MSCs (stroma) and the co-culture of stroma and tumorspheres (TME) cultured in the hydrogel. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Cells were centrifuged followed by the addition of fluorochrome-conjugated monoclonal antibodies for CD98 and CD44 (Miltenyi Biotec) according to the manufacturer's instructions and incubated at 4 °C in the dark for 12 min. After adding BSA, cells were centrifugated and resuspended in PBS and analyzed by flow cytometry in a FACSCanto II cytometer (BD Biosciences).

Techniques: Suspension, Cytometry, Membrane, Expressing, Activity Assay, Co-Culture Assay, Cell Culture, Proliferation Assay

Journal: Neurobiology of disease

Article Title: Identifying disulfidptosis-related biomarkers in epilepsy based on integrated bioinformatics and experimental analyses.

doi: 10.1016/j.nbd.2025.106789

Figure Lengend Snippet: Fig. 2. Identification of DE-DRMs in individuals with EP. (A) The heatmap of expression levels for nine DE-DRMs. (B) The expression levels of 11 DRMs were exhibited between Ctrl and EP groups in boxplots. (C) Correlation analysis of nine DE-DRMs. Red and green colors represent positive and negative correlations, respectively. (D) The PPI analysis of DE-DMRs showed that SLC3A2 interacted with SLC7A11, while NDUFS1 interacted with NDUFA11, NUBPL, and LRPPRC. Abbreviations: DE-DRMs, differentially expressed disulfidptosis-related molecules; EP, epilepsy; Ctrl, control; PPI, protein-protein interaction; SLC3A2, solute carrier family 3 member 2; SLC7A11, solute carrier family 7 member 11; NDUFS1, NADH:ubiquinone oxidoreductase core subunit S1; NDUFA11, NADH:ubiquinone oxidoreductase subunit A11; NUBPL, NUBP iron‑sulfur cluster assembly factor, mitochondrial; LRPPRC, leucine rich pentatricopeptide repeat containing. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The primary antibodies for western blot were as follows: anti-SLC3A2 antibody (Santa, sc-390,154, dilution: 1:500), anti-SLC7A11 antibody (Abcam, ab307601, dilution: 1:1000), anti-NDUFS1 antibody (Abcam, ab185733, dilution: 1:1000), anti-LRPPRC antibody (Abcam, ab259927, dilution: 1:1000), antiNDUFA11 antibody (Abclonal, A16239, dilution: 1:1000), and antiNUBPL antibody (Boster, A10634–1, dilution: 1:1000).

Techniques: Expressing, Control

Journal: Neurobiology of disease

Article Title: Identifying disulfidptosis-related biomarkers in epilepsy based on integrated bioinformatics and experimental analyses.

doi: 10.1016/j.nbd.2025.106789

Figure Lengend Snippet: Fig. 8. The expression of DE-DRMs in seizures models (in vitro and in vivo). (A) Constructing the in vitro seizures model. The amplitude and frequency of neuronal APs in the Mg2+-free group were significantly increased (n = 6 in each group; Independent sample t-test; #, P < 0.01). (B) Protein expression of nine DE-DRMs in the in vitro seizures model. The expression of GYS1, NDUFS1, OXSM, LRPPRC, NDUFA11, NUBPL, NCKAP1, and SLC3A2 were not significantly changed in the Mg2+-free group, while the expression of SLC7A11 was significantly increased in Mg2+-free group (n = 6 in each group; Independent sample t-test; #, P < 0.01). (C) Con structing the in vivo seizures model. No epileptoid discharges were observed in six rats of the Ctrl group, while significant epileptoid discharges were observed in six rats of the PTZ group (scale: Y-axis,50uV; X-axis, 0.5 s). (D) Protein expression of nine DE-DRMs in vivo models. The expressions of GYS1, NDUFS1, OXSM, LRPPRC, NDUFA11, NUBPL, NCKAP1, and SLC3A2 were not significantly changed in PTZ group, while the expression of SLC7A11 was significantly increased in PTZ group (n = 6 in each group; Independent sample t-test; #, P < 0.01). Abbreviations: DE-DRMs, differentially expressed disulfidptosis-related molecules; APs, action potentials; GYS1, glycogen synthase 1; SLC3A2, solute carrier family 3 member 2; SLC7A11, solute carrier family 7 member 11; NDUFS1, NADH:ubiquinone oxidoreductase core subunit S1; OXSM, 3-oxoacyl-ACP synthase, mitochondrial; LRPPRC, leucine rich pentatricopeptide repeat containing; NDUFA11, NADH:ubiquinone oxidoreductase subunit A11; NUBPL, NUBP iron‑sulfur cluster assembly factor, mitochondrial; NCKAP1, NCK associated protein 1; PTZ, pentylenetetrazol.

Article Snippet: The primary antibodies for western blot were as follows: anti-SLC3A2 antibody (Santa, sc-390,154, dilution: 1:500), anti-SLC7A11 antibody (Abcam, ab307601, dilution: 1:1000), anti-NDUFS1 antibody (Abcam, ab185733, dilution: 1:1000), anti-LRPPRC antibody (Abcam, ab259927, dilution: 1:1000), antiNDUFA11 antibody (Abclonal, A16239, dilution: 1:1000), and antiNUBPL antibody (Boster, A10634–1, dilution: 1:1000).

Techniques: Expressing, In Vitro, In Vivo

Journal: Neurobiology of disease

Article Title: Identifying disulfidptosis-related biomarkers in epilepsy based on integrated bioinformatics and experimental analyses.

doi: 10.1016/j.nbd.2025.106789

Figure Lengend Snippet: Fig. 9. Verifying the PPI in seizures models (in vivo and in vitro). (A-B) colocation analysis indicated that SLC7A11 colocalized with SLC3A2, NDUFS1 colocalized with LRPPRC, NDUFS1 colocalized with NUBPL, and NDUFS1 colocalized with NDUFA11 in both primary neurons and hippocampal tissue. (C–D) Pooled quan tification of protein immunoprecipitation showed a significant increment in the pull-down of SLC7A11 and a significant reduction in the pull-down of NDUFA11 in both the Mg2+-free group and PTZ group (n = 6 in each group; Independent sample t-test; #, P < 0.01). Abbreviations: PPI, protein-protein interaction; SLC3A2,solute carrier family 3 member 2; SLC7A11, solute carrier family 7 member 11; NDUFS1, NADH:ubiquinone oxidoreductase core subunit S1; LRPPRC, leucine rich pentatricopeptide repeat containing; NUBPL, NUBP iron‑sulfur cluster assembly factor, mitochondrial; NDUFA11, NADH:ubiquinone oxidoreductase subunit A11; PTZ, pentylenetetrazol.

Article Snippet: The primary antibodies for western blot were as follows: anti-SLC3A2 antibody (Santa, sc-390,154, dilution: 1:500), anti-SLC7A11 antibody (Abcam, ab307601, dilution: 1:1000), anti-NDUFS1 antibody (Abcam, ab185733, dilution: 1:1000), anti-LRPPRC antibody (Abcam, ab259927, dilution: 1:1000), antiNDUFA11 antibody (Abclonal, A16239, dilution: 1:1000), and antiNUBPL antibody (Boster, A10634–1, dilution: 1:1000).

Techniques: In Vivo, In Vitro, Immunoprecipitation

Journal: Frontiers in Oncology

Article Title: Decrease of Intracellular Glutamine by STF-62247 Results in the Accumulation of Lipid Droplets in von Hippel-Lindau Deficient Cells

doi: 10.3389/fonc.2022.841054

Figure Lengend Snippet: Effects of STF-62247 on glutamine transporters. (A, B) mRNA relative expression, measured by RT-qPCR, of glutamine transporters SLC1A5, SLC7A5 and SLC3A2 influenced by (A) VHL status and (B) STF-62247. RCC4 VHL- and VHL+ were treated with 1.25 µM STF-62447 for 24 hr. Results for treated cells are compared to control cells (represented by the dotted line at 1). Results are presented as means and SEM of three independent experiments. (C) Glutamine transporters, SLC1A5, SL7A5 and SLC3A2, protein levels in RCC4 VHL- and VHL+ treated to STF-62247 for 24 and 48 hr. (D) Intracellular leucine was measured by LC-MS in RCC4 VHL- and VHL+ treated with 1.25 µM STF-62247 for 24 hr (N=6). Student’s t -tests were performed to compare results between VHL- and VHL+ cells (A) or between controls and treated cells (B, D) (*p < 0.05, **p < 0.01, ***p < 0.001). (E) Cell viability was evaluated by XTT assay in RCC4 VHL- and VHL+ cells. SLC1A5 and SLC7A5 inhibitors, GPNA and BCH respectively, were tested alone (concentrations from 0 to 1 mM) or combined with 1.25 µM STF-62247 (N=3). Two-Way ANOVA followed by Tukey’s test was performed to assess statistically significant results. Comparison between CTL and STF-62247 conditions for each concentration of inhibitor (GPNA or BCH) are denoted with the following statistical marks *p < 0.05, **p < 0.01 or ***p < 0.001. Comparison of each inhibitor concentrations (x-axis) are made with their respective control (0 mM columns, with or without STF-62247) and statistical significances are denoted by # p < 0.05, ## p < 0.01 or ### p < 0.001.

Article Snippet: Membranes were blocked using 5% skim milk diluted in a solution of 0.075% PBS-Tween (PBS-T) and incubated overnight in 3% BSA with specific primary antibodies against VHL, HIF-1α and HIF-2α (Cell signaling #68547, 14179, 59973), SLC1A5 and β-actin (Santa Cruz Biotechnologies sc-99002 and sc-47778), SLC7A5 (Medical & Biological Laboratories #BMP011), SLC3A2 (Aviva Systems Biology #OAAB00158), ASNS (Signalway #32909), SCD1 (Applied Biological Materials #ABM-G076), PLIN2 (Proteintech #15294-1-AP) and CPT1A (Abcam #ab220789).

Techniques: Expressing, Quantitative RT-PCR, Control, Liquid Chromatography with Mass Spectroscopy, XTT Assay, Comparison, Concentration Assay

Journal: Molecular Pharmaceutics

Article Title: Structural Features Affecting the Interactions and Transportability of LAT1-Targeted Phenylalanine Drug Conjugates

doi: 10.1021/acs.molpharmaceut.2c00594

Figure Lengend Snippet: Confocal microscopy images after immunofluorescence staining of HEK-MOCK cells (left) and HEK-hLAT1 cells (middle and right), showing LAT1 localization in the cell membrane (LAT1 = red, nuclei of the cells = blue, 4F2hc = green, overlapping of LAT1/4F2hc = yellow/orange).

Article Snippet: The cells were incubated with primary antibody dilutions 1:50 at RT for 1.5 h (LAT1#5347 for LAT1, Cell Signaling and AM33318PU-T for 4F2hc, Origene).

Techniques: Confocal Microscopy, Immunofluorescence, Staining