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: 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: 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: 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: Bone & Joint Research

Article Title: The role of EDIL3 in maintaining cartilage extracellular matrix and inhibiting osteoarthritis development

doi: 10.1302/2046-3758.1212.BJR-2023-0087.R1

Figure Lengend Snippet: EGF-like repeats and discoidin I-like domains-containing protein 3 (EDIL3) has beneficial effects on cartilage maintenance in mice with osteoarthritis (OA). a) Schematic representation of the timeline of the antibody drug or recombinant protein (EDIL3 or CD98) treatment during 16 to 21 weeks of age. STR/ort mice were injected weekly through the tail vein with phosphate-buffered saline (PBS, vehicle) combined with either the antibody or the protein for six consecutive weeks. Mice were killed at 22 weeks. This was followed by paraffin tissue sections and Safranin O and immunofluorescence (IF) staining in knee articular cartilage. b) Representative images include articular cartilage in the tibial plateau knee joints. Arrows indicate representative matrix-producing (red) and matrix-non-producing (black) chondrocytes. c) The total chondrocyte number in the articular cartilage was quantified. EDIL3 protein treatment increased the number of chondrocytes. d) The number and percentage of matrix-non-producing chondrocytes (MNCs) in the articular cartilage were quantified. EDIL3 antibody treatment increased the number of MNCs. e) EDIL3 protein treatments decreased the Osteoarthritis Research Society International (OARSI) score in the LFC. f) Representative images of IF staining (green) in whole articular cartilage obtained from STR/ort mice for the EDIL3; 4',6-diamidino-2-phenylindole (DAPI) (blue) stained nuclei; orange dashed lines define the cartilage region. EDIL3 antibody treatments decreased the EDIL3 contents in the cartilage region. g) The fluorescence of EDIL3 was quantified in the whole articular cartilage region. EDIL3 antibody significantly decreased EDIL3 expression. h) Representative images of IF staining (green) in whole articular cartilage obtained from STR/ort mice for the indicated OA markers; DAPI (blue) stained nuclei; orange dashed lines define the cartilage region. i) Fluorescence was quantified in the whole articular cartilage region. Data are presented as means and standard errors in . Data are presented as mean and maximum with 95% confidence intervals in . CD98 protein significantly increased SOX9 expression, and EDIL3 protein significantly reduced aggrecan fragments. LTP, lateral tibial plateau; MFC, medial femoral condyle; MMP, matrix metalloproteinase; MTP, medial tibial plateau. Data presented in Figures 3c to 3e were analyzed using one-way analysis of variance (ANOVA) followed by Tukey’s multiple comparison test for selected pairs of groups for multiple comparisons. *p < 0.05, **p < 0.01, ***p < 0.001. Data in Figures 3g to 3i were analyzed using one-way ANOVA followed by Dunnett’s multiple comparison test. Isotype Ab, immunoglobulin G (IgG)1 antibody.

Article Snippet: The STR/ort mice were administered tail-vein injections of 1 μg/mouse anti-EDIL3 IgG1 (MABS1976; MilliporeSigma), 2 μg/mouse recombinant EDIL3 protein (6046-ED; R&D Systems), 2 μg/mouse CD98 protein (50813-M07H; Sino Biological, China), 1 μg/mouse isotype-matched control IgG1 monoclonal antibody (mAb) (mabg1-ctrlm; InvivoGen, USA), or vehicle PBS solution.

Techniques: Recombinant, Injection, Saline, Immunofluorescence, Staining, Fluorescence, Expressing, Comparison

Journal: Bone & Joint Research

Article Title: The role of EDIL3 in maintaining cartilage extracellular matrix and inhibiting osteoarthritis development

doi: 10.1302/2046-3758.1212.BJR-2023-0087.R1

Figure Lengend Snippet: EGF-like repeats and discoidin I-like domains-containing protein 3 (EDIL3) treatment prevents subchondral bone plate thickness (PI.Th) and epiphyseal trabecular mineralization. a) Representative cross and horizontal sections of knee joints obtained from STR/ort mice. The 3D reconstruction of a proximal tibia with a plane indicates the location from which the cross and horizontal sections were obtained. The vehicle control mice exhibited uneven trabecular bone distribution; moreover, EDIL3 antibody treatment was observed to worsen this phenomenon. However, EDIL3 and CD98 protein prevented subchondral bone mineralization. b) Trabecular bone morphometric parameters were calculated using micro-CT analysis. The medial and lateral subchondral bone plate and underlying epiphyseal trabecular bone were analyzed separately. Epiphysis was manually selected as representative of subchondral bone. The epiphyseal trabeculae were split from the subchondral bone plate, and trabecular bone morphometric parameters were calculated. EDIL3 protein treatment prevented osteoarthritis (OA)-associated reduction in subchondral bone total porosity (Po(tot)). EDIL3 protein treatment also decreased the subchondral PI.Th, bone volume (BV/TV), and trabecular parameters (trabecular thickness (Tb.Th), trabecular number (Tb.N), and trabecular spacing (Tb.Sp)). Analyses were conducted with two-way analysis of variance (ANOVA) followed by Tukey’s multiple comparisons test. Data are presented as means and standard deviations. *p < 0.05, **p < 0.01; analyzed using a two-way analysis of variance followed by Tukey’s multiple comparison test.

Article Snippet: The STR/ort mice were administered tail-vein injections of 1 μg/mouse anti-EDIL3 IgG1 (MABS1976; MilliporeSigma), 2 μg/mouse recombinant EDIL3 protein (6046-ED; R&D Systems), 2 μg/mouse CD98 protein (50813-M07H; Sino Biological, China), 1 μg/mouse isotype-matched control IgG1 monoclonal antibody (mAb) (mabg1-ctrlm; InvivoGen, USA), or vehicle PBS solution.

Techniques: Control, Micro-CT, Comparison

Journal: Bone & Joint Research

Article Title: The role of EDIL3 in maintaining cartilage extracellular matrix and inhibiting osteoarthritis development

doi: 10.1302/2046-3758.1212.BJR-2023-0087.R1

Figure Lengend Snippet: EGF-like repeats and discoidin I-like domains-containing protein 3 (EDIL3) inhibition promotes pro-inflammatory cytokine production in STR/ort mice. a) The variable expressions of 48 cytokines among vehicle control, isotype IgG1, EDIL3 IgG1, recombinant EDIL3 protein, and CD98 protein groups were identified by comparing the expression pattern in the serum of STR/ort mice and those with vehicle control group by heat-map analysis. Arrows indicate representative anti-inflammatory cytokines (red) and pro-inflammatory cytokines (green). b) Quantitation of each cytokine demonstrated serum samples from different groups using Luminex xMAP technology. Six anti-inflammatory cytokines and 20 pro-inflammatory cytokines were identified with the most significant difference between the five groups. c) Notably, many pro-inflammatory cytokines in serum increase with ageing, including monocyte chemotactic protein-3 (MCP-3), RANTES, interleukin (IL)-17A, IL-22, and GRO-alpha. EDIL3 antibody significantly promoted the increase of these pro-inflammatory cytokines. Analyses were conducted with one-way analysis of variance (ANOVA) followed by Tukey’s multiple comparisons test for selected pairs of groups. Data are presented as means and standard errors. *p < 0.05, **p < 0.01, ***p < 0.001. Data presented in Figure 5c were analyzed using one-way analysis of variance followed by Tukey’s multiple comparison test for selected pairs of groups for multiple comparisons. IgG, immunoglobulin G.

Article Snippet: The STR/ort mice were administered tail-vein injections of 1 μg/mouse anti-EDIL3 IgG1 (MABS1976; MilliporeSigma), 2 μg/mouse recombinant EDIL3 protein (6046-ED; R&D Systems), 2 μg/mouse CD98 protein (50813-M07H; Sino Biological, China), 1 μg/mouse isotype-matched control IgG1 monoclonal antibody (mAb) (mabg1-ctrlm; InvivoGen, USA), or vehicle PBS solution.

Techniques: Inhibition, Control, Recombinant, Expressing, Quantitation Assay, Luminex, Comparison