Journal: Diabetes

Article Title: Adiponectin reduces glomerular endothelial glycocalyx disruption and restores glomerular barrier function in a mouse model of type 2 diabetes.

doi: 10.2337/db23-0455

Figure Lengend Snippet: Figure 3—Adiponectin ameliorates TNFa-induced glycocalyx shedding. GEnCs were treated with or without 10 ng/mL TNF-a for 2 h in the presence or absence of 2.5 mg/mL gAd. A) qPCR analysis of SDC4 mRNA levels, normalized to GAPDH, are shown. Data are plotted as the mean 2-(DCT) (gene of interest/housekeeping gene) of each triplicate with the mean. B) SDC4 protein expression was quantified by ELISA in the medium of treated cells. C) Sulfated GAGs were quantified in the medium of treated cells by Alcian blue colorimetric assay. MMP2 (D) and MMP9 (E) mRNA levels in CiGEnCs treated with TNF-a (10 ng/mL) and/or gAd (2.5 mg/mL). A–E) One-way ANOVA (n = 5); *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 by post hoc analysis (Bonferroni). MMP2 was knocked down using three shRNA dif- ferent sequences in GEnCs (v33, v47, v49) or not, using scrambled controls. F) qPCR data analysis highlighting the decreased expression of MMP2 mRNA in CiGEnCs by all three different shRNA sequences. Data are plotted as the mean comparative cycle threshold of each triplicate. GAPDH used as the housekeeping gene control, n = 4; one-way ANOVA, ****P < 0.0001 post hoc analysis (Bonferroni). Gi) Representative Western Blot demonstrating the protein knockdown of MMP2 by shRNA clone v47. Gii) Densitometry showed significant knockdown of MMP2 protein expression by clone v47. Data are normalized to the b-actin loading control. Dots represent mean ± SEM; one-way ANOVA, ***P < 0.001 by post hoc analysis (Bonferroni). H) SDC4 mRNA levels in MMP2 knockdown CiGEnCs treated with TNF-a and/or gAd. Data are plotted as the mean comparative cycle threshold of each triplicate. GAPDH was used as the housekeeping gene control, n = 5; one-way ANOVA, ***P < 0.001 post hoc analysis (Bonferroni).

Article Snippet: Cellular levels of SDC4 were quantified using a SDC4 ELISA (R&D Systems; catalog DY2918) per the manufacturer’s instructions.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Colorimetric Assay, shRNA, Control, Western Blot, Knockdown

Journal: Diabetes

Article Title: Adiponectin reduces glomerular endothelial glycocalyx disruption and restores glomerular barrier function in a mouse model of type 2 diabetes.

doi: 10.2337/db23-0455

Figure Lengend Snippet: Figure 4—Adiponectin signals to glomeruli directly and prevents the mRNA upregulation of glycocalyx-related genes in db/db glomeruli. Hu- man (A) and mouse (B) glomeruli were isolated and treated ex vivo with 2.5 mg/mL gAd. Representative Western Blots demonstrating gAd ef- fects at 30 min in human (Ai) and mouse (Bi) ex vivo glomeruli on AMPK phosphorylation (p-). Aii and Bii) Densitometry confirmed significant phosphorylation of AMPK in response to gAd after 30 min in human and mouse glomeruli. Densitometry was performed on blots from inde- pendent repeats (n = 4) showing levels of protein of interest normalized to the b-actin loading control. Dots represent mean ± SEM; one-way ANOVA; *P < 0.05, by post hoc analysis (Bonferroni). C–F) qPCR analysis graph representing the mRNA expression of TNF (C), SDC4 (D), MMP2 (E), and MMP9 (F) in ex vivo (wild type or db/db) sieved glomeruli treated with or without adiponectin. Data are plotted as the mean comparative cycle threshold of each triplicate, n = 5; one-way ANOVA; *P < 0.05, **P < 0.01, ****P < 0.0001 by post hoc analysis (Bonferroni).

Article Snippet: Cellular levels of SDC4 were quantified using a SDC4 ELISA (R&D Systems; catalog DY2918) per the manufacturer’s instructions.

Techniques: Isolation, Ex Vivo, Western Blot, Phospho-proteomics, Control, Expressing

Journal: Oncogene

Article Title: LAMP5 modulates IRF4 stability and nuclear transport: a critical mechanism in myeloma progression and therapy

doi: 10.1038/s41388-025-03513-x

Figure Lengend Snippet: A , B LAMP5 mRNA expression in myeloma and other indicated cancer cell lines or other tissues, according to the Cancer Cell Line Encyclopedia (CCLE) dataset. C LAMP5 mRNA levels in plasma cells from myeloma patients ( n = 559) compared to normal plasma cells from healthy donors ( n = 22) (GEO: GSE2658 and GSE5900 ) (left). LAMP5 mRNA levels in plasma cells from myeloma patients ( n = 73) compared to normal plasma cells from healthy donors ( n = 14) (GEO: GSE6477 ) (right). P value was determined by unpaired two-tailed t test. D Quantitative real-time PCR analysis shows LAMP5 expression in normal plasma cells (Healthy, n = 10), malignant plasma cells (Pt MM, n = 20), and human myeloma cell lines (MM cell lines, n = 6). GAPDH served as loading control. P values were determined using one-way ANOVA. E Representative two patients of UMAP plot of CD138 (left) and LAMP5 (right) expression in scRNA-seq data ( GSE161801 ). F Immunofluorescent staining of healthy of myeloma patients bone marrow samples with DAPI and antibodies against LAMP5 and CD138 ( n = 5). Scale bar, 10 μm. G Overall survival (OS) of patient’s myeloma cells with high LAMP5 (High, n = 200) and low LAMP5 (Low, n = 200) expression in MMRF CoMMpass study IA15.

Article Snippet: The CD138 antibody (#AF2780) was purchased from R&D Systems.

Techniques: Expressing, Clinical Proteomics, Two Tailed Test, Real-time Polymerase Chain Reaction, Control, Staining

Journal: Oncogene

Article Title: LAMP5 modulates IRF4 stability and nuclear transport: a critical mechanism in myeloma progression and therapy

doi: 10.1038/s41388-025-03513-x

Figure Lengend Snippet: A Analysis of immunoprecipitates of HA-tagged LAMP5 in IM-9 cells with mass spectrometry. The proteins identified are indicated. B Co-immunoprecipitation of LAMP5 with IRF4 in ARP-1 or IM-9 cells. C Co-immunoprecipitation of LAMP5 with IRF4 in HEK293T cells co-transfected with LAMP5 and IRF4 plasmids. D Pull-down of HA-LAMP5 with IRF4 in HEK293T cells. E Immunofluorescent staining of ARP-1 or IM-9 cells with DAPI and antibodies against LAMP5 and IRF4. Scale bar, 10 μm. F Immunofluorescent staining of myeloma patients bone marrow samples with DAPI and antibodies against CD138, LAMP5 and IRF4 ( n = 5). Scale bar, 10 μm. G Schematic of the truncations including ΔDBD and ΔIAD fragments. H Western blotting showing different truncations of IRF4 (ΔDBD and ΔIAD) in HEK293T cells. I Pull-down of LAMP5 with different truncations of IRF4 (ΔDBD and ΔIAD) in HEK293T cells. J – M The expression of IRF4 or c-MYC in myeloma cells with reduced (sh LAMP5 ) or increased ( LAMP5 ) LAMP5 expression. Nontargeted shRNA (sh Ctrl ) or control vector ( Vec ) served as control. N , O The expression of LDHA or KLF2 in myeloma cells with reduced (sh LAMP5 ) LAMP5 expression. Nontargeted shRNA (sh Ctrl ) served as control. P , Q Immunofluorescent staining of ARP-1 or IM-9 cells (sh Ctrl and sh LAMP5 ) with DAPI and antibody against LAMP5 and IRF4. Scale bar, 10 μm. R Immunofluorescent staining of tumor tissues in Fig. with DAPI and antibodies against LAMP5 and IRF4. Scale bar, 20 μm. S Correlation coefficient of the mRNA levels of LAMP5 and mRNA levels of IRF4 or c-MYC in myeloma patients ( n = 559). The correlations were evaluated using Pearson coefficient. r, correlation coefficient. B – D , H , I , and M are representative of two independent experiments. E , J –L, and N – R are representative of three independent experiments. Data are averages ±SD. P value was determined by Pearson coefficient. J , K , L , N , O P values were determined using one-way ANOVA.

Article Snippet: The CD138 antibody (#AF2780) was purchased from R&D Systems.

Techniques: Mass Spectrometry, Immunoprecipitation, Transfection, Staining, Western Blot, Expressing, shRNA, Control, Plasmid Preparation

Journal: Oncogene

Article Title: LAMP5 modulates IRF4 stability and nuclear transport: a critical mechanism in myeloma progression and therapy

doi: 10.1038/s41388-025-03513-x

Figure Lengend Snippet: A Workflow of high-throughput virtual screening (HTVS) for LAMP5 inhibitors. B Docking scores of the top 4 candidates obtained from HTVS. C Chemical structure and in silico docking of pyrazofurin into the active pocket of human LAMP5 protein. D Proliferation of myeloma cells (ARP-1 or IM-9) treated with or without pyrazofurin (Pyr.) (0.1 or 1 μM) over time. PBS buffer treated group served as control. E Antiproliferative activities of pyrazofurin were evaluated on human myeloma cell lines ARP-1 or IM-9. F Co-immunoprecipitation of LAMP5 with IRF4 in ARP-1 or IM-9 cells treated with or without pyrazofurin (1 μM). G Co-immunoprecipitation of LAMP5 with IRF4 in HEK293T cells co-transfected with HA-LAMP5 and IRF4 plasmids. H IRF4 protein levels in myeloma cells treated with 1 μM pyrazofurin and 2.5 μM MG-132 or 2.5 μM leupeptin for 7 h. 6-week-old male C57BL/6 J mice were intravenously injected vk12598 cells ( n = 9 mice/group), followed by intraperitoneal administration of pyrazofurin (5 mg/kg bodyweight) three times per week for a duration of two weeks, beginning two weeks after cell injection. Shown are the experimental schematic ( I ). J ELISA analysis shown the concentrations of IgG2b in mouse sera. K Immunofluorescent staining of I mentioned spleen tumor cells of vk12598 mice with DAPI and antibodies against CD138 or IRF4. Scale bar, 40 μm. L Photographic images of tumors in NSG mice after implantation of ARP-1 or IM-9 cells ( n = 3 mice/group), and intraperitoneal administration with or without pyrazofurin (5 mg/kg bodyweight). Shown are time course of tumor volume ( M ). N , O Immunofluorescent staining of Ki67 or IRF4 expression in tumor tissues. D and E are representative of three independent experiments. F – H are representative of two independent experiments. Data are averages ±SD. J P value was determined by unpaired two-tailed t test; D , M P values were determined using two-way ANOVA.

Article Snippet: The CD138 antibody (#AF2780) was purchased from R&D Systems.

Techniques: High Throughput Screening Assay, In Silico, Control, Immunoprecipitation, Transfection, Injection, Enzyme-linked Immunosorbent Assay, Staining, Expressing, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Syndecan-4 Is a Key Facilitator of the SARS-CoV-2 Delta Variant’s Superior Transmission

doi: 10.3390/ijms23020796

Figure Lengend Snippet: Cellular uptake of the WT spike and its Delta variant into WT K562 cells and SDC1, 4 transfectants. WT K562 cells, SDC1 and SDC4 transfectants were treated with the WT spike and its Delta variant. The cellular uptake was then analyzed with imaging flow cytometry by using fluorescently labeled specific antibodies against the His-tag of the recombinant spikes. ( A , B ) Representative flow cytometry histograms and cellular images show the intracellular fluorescence of WT K562 cells, SDC1 and SDC4 transfectants treated with either of the spike proteins. Scale bar = 20 μm. ( C ) Detected fluorescence intensities normalized to WT-spike-treated K562 cells as standards. The bars represent the mean + SEM of six independent experiments (data are represented as dots). Statistical significance vs. standards was assessed with ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001. ( D ) Detected fluorescence intensities normalized to WT-spike-treated cells as standards. The bars represent the mean + SEM of six independent experiments (data are represented as dots). Statistical significance vs. standards was assessed with ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: The supernatant was then transferred to new tubes and combined with 5 μg human SDC4 antibody (RnD System, UK; cat. no. AF2918).

Techniques: Variant Assay, Imaging, Flow Cytometry, Labeling, Recombinant, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Syndecan-4 Is a Key Facilitator of the SARS-CoV-2 Delta Variant’s Superior Transmission

doi: 10.3390/ijms23020796

Figure Lengend Snippet: PSV-mediated gene transduction into WT K562 cells, SDC1 and SDC4 transfectants. The cells were incubated with SARS-CoV-2 PSV-RFP (either the WT or the Delta variant). RFP expression was analyzed 72 h later with imaging flow cytometry. ( A ) Representative flow cytometry histograms showing RFP fluorescence of WT K562 cells, SDC1 and SDC4 transfectants incubated with either the WT or the Delta SARS-CoV-2 PSV. ( B ) Cellular images of SARS-CoV-2 PSV-treated WT K562 cells and SDC transfectants as detected with imaging flow cytometry. Scale bar = 20 μm. ( C ) Detected cellular RFP intensities normalized to WT PSV-treated WT K562 cells as standards. The bars represent the mean + SEM of three independent experiments (data are represented as dots). Statistical significance vs. standards was assessed with ANOVA. * p < 0.05; ** p < 0.01. ( D ) Detected RFP intensities normalized to WT PSV-treated cells as standards. The bars represent the mean ± SEM of three independent experiments (data are represented as dots). Statistical significance vs. standards was assessed with ANOVA. * p < 0.05; *** p < 0.001.

Article Snippet: The supernatant was then transferred to new tubes and combined with 5 μg human SDC4 antibody (RnD System, UK; cat. no. AF2918).

Techniques: Transduction, Incubation, Variant Assay, Expressing, Imaging, Flow Cytometry, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Syndecan-4 Is a Key Facilitator of the SARS-CoV-2 Delta Variant’s Superior Transmission

doi: 10.3390/ijms23020796

Figure Lengend Snippet: Cellular uptake of the WT and Delta spike proteins into SDC4 deletion mutants. WT SDC4 transfectants and CBD, HSA and Si4 mutants (all tagged with GFP) were treated with the Delta spike. The cellular uptake was then analyzed with imaging flow cytometry by using fluorescently labeled specific antibodies against the His-tag of the recombinant spike proteins. ( A ) Representative flow cytometry histograms showing the intracellular fluorescence of WT SDC4 transfectants, CBD, HSA and Si4 mutants treated with the Delta spike protein (along with specific fluorescent antibodies). Scale bar = 20 μm. ( B ) Detected fluorescence intensities were normalized Delta spike-treated WT SDC4 transfectants as standards. The bars represent the mean ± SEM of four independent experiments (experimental data are represented as dots). Statistical significance vs. standards was assessed with ANOVA. *** p < 0.001. ( C ) Cellular images showing the intracellular fluorescence of WT SDC4 and CBD, HSA and Si4 mutants treated with the Delta spike protein. Scale bar = 20 μm. The indicated BDS values of SARS-CoV-2 and SDCs represent the mean + SEM of four independent experiments.

Article Snippet: The supernatant was then transferred to new tubes and combined with 5 μg human SDC4 antibody (RnD System, UK; cat. no. AF2918).

Techniques: Imaging, Flow Cytometry, Labeling, Recombinant, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Syndecan-4 Is a Key Facilitator of the SARS-CoV-2 Delta Variant’s Superior Transmission

doi: 10.3390/ijms23020796

Figure Lengend Snippet: Internalization of the WT and Delta spike into Calu-3 cells. Calu-3 cells were treated with the WT spike and its Delta variant. The cellular uptake was then analyzed with imaging flow cytometry by using fluorescently labeled specific antibodies against the His-tag of the recombinant spike proteins. ( A ) Representative flow cytometry histograms and cellular images show the intracellular fluorescence of Calu-3 cells treated with either the WT or Delta spike. ( B ) Detected intracellular fluorescence levels normalized to those treated with the WT spike as standards. The bars represent the mean + SEM of three independent experiments. Statistical significance vs. 293T expression levels was assessed with ANOVA. * p < 0.05. ( C ) SDS-PAGE showing the WT or Delta spike immunoprecipitated with SDC4 antibodies from extracts of untreated control or spike-treated Calu-3 cells. Lanes 1–2: 0.5 ug of WT and Delta spike proteins; Lanes 3–4: immunoprecipitate of WT or Delta spike-treated Calu-3 cells; Lane 5: immunoprecipitate of untreated Calu-3. Standard protein size markers are indicated on the left. The signal of spike proteins was detected with UVITEC Alliance Q9 Advanced Imager, and the intensity of bands were analyzed with the NineAlliance © software. ( D ) Detected band intensities were normalized to WT spike-treated Calu-3 cells as standard. The bars represent the mean + SEM of three independent experiments. Statistical significance vs. standards was assessed with ANOVA. * p < 0.05.

Article Snippet: The supernatant was then transferred to new tubes and combined with 5 μg human SDC4 antibody (RnD System, UK; cat. no. AF2918).

Techniques: Variant Assay, Imaging, Flow Cytometry, Labeling, Recombinant, Fluorescence, Expressing, SDS Page, Immunoprecipitation, Control, Software

Journal: International Journal of Molecular Sciences

Article Title: Syndecan-4 Is a Key Facilitator of the SARS-CoV-2 Delta Variant’s Superior Transmission

doi: 10.3390/ijms23020796

Figure Lengend Snippet: Effect of SDC4 knockdown (KD) or GAG inhibition on Delta spike internalization into Calu-3 cells. SDC4 KD in Calu-3 cells was performed using a lentiviral vector specific to human SDC4. ( A ) SDC4 expression levels were measured with imaging flow cytometry, as shown by the representative histograms and cellular images. Detected SDC4 levels of KD cells were normalized to WT Calu-3 cells as standards. The bars represent the mean + SEM of three independent experiments. Statistical significance vs. standards was assessed with ANOVA. * p < 0.05. ( B,C ) SDC4 KD and WT Calu-3 cells were treated with either the Delta spike protein ( B ) or the Delta PSV ( C ). For GAG inhibition, some of the WT Calu-3 cells were preincubated with heparin (200 ug/mL, 30 min before Delta treatment). Representative flow cytometry histograms and cellular images show the intracellular fluorescence of WT or SDC4 KD Calu-3 treated with Delta spike protein or PSV in the presence or absence of heparin. Scale bar = 20 μm. The effect of SDC4 KD or heparin treatment was expressed as percent inhibition, calculated with the following formula: ((X − Y)/X) × 100, where X is the fluorescence intensity obtained on controls treated with the Delta spike or Delta PSV without SDC4 KD or heparin, and Y is the fluorescence intensity obtained on cells receiving Delta spike or Delta PSV treatment, along with SDC4 KD or heparin. The bars represent the mean + SEM of three independent experiments. Statistical significance vs. controls was assessed with ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: The supernatant was then transferred to new tubes and combined with 5 μg human SDC4 antibody (RnD System, UK; cat. no. AF2918).

Techniques: Knockdown, Inhibition, Plasmid Preparation, Expressing, Imaging, Flow Cytometry, Fluorescence

Journal: Journal of Cellular and Molecular Medicine

Article Title: Cell surface‐anchored syndecan‐1 ameliorates intestinal inflammation and neutrophil transmigration in ulcerative colitis

doi: 10.1111/jcmm.12934

Figure Lengend Snippet: Levels of shed SDC 1 and P65, secretion of TNF ‐α and IL ‐1β, as well as circulating neutrophil rate in ulcerative colitis patients. ( A ) Representative immunostaining of SDC 1, P65 and CD 15 in intestinal mucosa with overview (100×) and a magnification (400×). The right lane: haematoxylin and eosin staining. ( B ) Levels of shed SDC 1 in serum was detected by ELISA ; n = 20. ( C ) Levels of secreted TNF ‐α ( n = 20) and IL ‐1β ( n = 20) in serum were detected by ELISA ; Circulating CRP was significantly higher in UC patients at active stage, n = 20. ( D ) Circulating white blood cells and neutrophils were significantly higher in UC patients at active stage; n = 20. ( E ) Shed SDC 1 in serum was positively correlated with circulating neutrophil rate ( R = 0.634, P < 0.01). n = 20. * P < 0.05, ** P < 0.01.

Article Snippet: Human Sdc1 ELISA kit was purchased from Elabscience Biotechnology Co., Ltd (Cat. E‐EL‐H1298; Wuhan, China).

Techniques: Immunostaining, Staining, Enzyme-linked Immunosorbent Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Cell surface‐anchored syndecan‐1 ameliorates intestinal inflammation and neutrophil transmigration in ulcerative colitis

doi: 10.1111/jcmm.12934

Figure Lengend Snippet: Shed SDC 1 and inflammatory responses induced by PMA in HT 29 cells. PMA was used to induce SDC 1 shedding in HT 29 cells. ( A ) Levels of Sdc1 in six intestinal epithelial cells were detected by western blot. GAPDH was used as the loading control. ( B ) Level of cell surface SDC 1 was detected by immunofluorescence. SDC 1 (red), cell nucleus (blue), original magnifications: 40×. ( C ) Levels of shed SDC 1 in the cell culture supernatant were detected by ELISA (upper) and dot blot (lower). ( D ) Levels of secreted TNF ‐α, IL ‐1β, IL ‐6 and IL ‐8 in the cell culture supernatant were detected by PCR . ( E ) Levels of P65 and phosphorylated P65 were detected by Western blot. ( F ) Secretion of CXCL ‐1 was detected by ELISA . ( G ) Neutrophils were isolated from human venous blood, and their transmigration was measured with use of transwell inserts. Crystal violet staining was used to observe the cell quantity change. Values represent mean ± S.E.M ( n = 3), * P < 0.05, ** P < 0.01, based on t ‐test.

Article Snippet: Human Sdc1 ELISA kit was purchased from Elabscience Biotechnology Co., Ltd (Cat. E‐EL‐H1298; Wuhan, China).

Techniques: Western Blot, Control, Immunofluorescence, Cell Culture, Enzyme-linked Immunosorbent Assay, Dot Blot, Isolation, Transmigration Assay, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: Cell surface‐anchored syndecan‐1 ameliorates intestinal inflammation and neutrophil transmigration in ulcerative colitis

doi: 10.1111/jcmm.12934

Figure Lengend Snippet: Cellular inflammatory responses induced by LPS was inhibited by cell surface‐anchored SDC 1 in Caco‐2 cells. SDC 1 shedding is blocked in Caco‐2 cells transfected with mut‐ SDC 1 plasmid and LPS was used to induce inflammatory responses. ( A ) Protein (upper) and mRNA (lower) levels of cell surface SDC 1 were measured by western blot and PCR respectively. GAPDH was used as the loading control. ( B ) Levels of shed SDC 1 in the cell culture supernatant were detected by ELISA (left upper) and dot blot (left lower). Protein level of cell surface SDC 1 upon LPS stimulation was measured by immunofluorescence (right). SDC 1 (red), cell nucleus (blue), original magnifications: 400×. ( C ) Cell surface‐anchored SDC 1 down‐regulated secretion of TNF ‐α, IL ‐1β, IL ‐6 and IL ‐8 in the cell culture supernatant detected by PCR . ( D ) Cell surface‐anchored SDC 1 inactivated NF ‐κB pathway detected by Western blot. ( E ) Cell surface‐anchored SDC 1 alleviated impaired viability induced by LPS , but had no significant effect on cell proliferation. Values represent mean ± S.E.M ( n = 3) and were analysed by Duncan's multiple range test for multiple comparison in anova (** P < 0.01. # significance between wt‐ SDC 1 and mut‐ SDC 1, ## P < 0.01).

Article Snippet: Human Sdc1 ELISA kit was purchased from Elabscience Biotechnology Co., Ltd (Cat. E‐EL‐H1298; Wuhan, China).

Techniques: Transfection, Plasmid Preparation, Western Blot, Control, Cell Culture, Enzyme-linked Immunosorbent Assay, Dot Blot, Immunofluorescence, Comparison

Journal: Journal of Cellular and Molecular Medicine

Article Title: Cell surface‐anchored syndecan‐1 ameliorates intestinal inflammation and neutrophil transmigration in ulcerative colitis

doi: 10.1111/jcmm.12934

Figure Lengend Snippet: Secreted CXCL ‐1 induced by LPS and sequent neutrophil transmigration was inhibited by cell surface‐anchored SDC 1 in Caco‐2 cells. LPS was use to induce secretions of CXCL ‐1, and expression of CXCL ‐1 was assessed by ELISA . ( A ) The level of secreted CXCL ‐1 was down‐regulated by Sdc1. ( B and C ) Cell culture supernatants containing high concentrations of CXCL ‐1 were used to induce migration of human neutrophils. CXCL ‐1 secretion promoted migration of neutrophils; this promoting effect was diminished when cells were transfected with mut‐ SDC 1. The transmigration was measured with use of transwell inserts, crystal violet staining was used to observe the cell quantity change. Values represent mean ± S.E.M ( n = 3) and were analysed by Duncan's multiple range test for multiple comparison in anova (* P < 0.05, ** P < 0.01. #significance between wt‐ SDC 1 and mut‐ SDC 1, # P < 0.05).

Article Snippet: Human Sdc1 ELISA kit was purchased from Elabscience Biotechnology Co., Ltd (Cat. E‐EL‐H1298; Wuhan, China).

Techniques: Transmigration Assay, Expressing, Enzyme-linked Immunosorbent Assay, Cell Culture, Migration, Transfection, Staining, Comparison