Journal: Frontiers in Physiology

Article Title: Dual roles of syndecan-4 in regulating chicken fibrosis in vitro

doi: 10.3389/fphys.2026.1782914

Figure Lengend Snippet: Overexpression of SDC4 in SL-29 chicken fibroblast cells. (A) Chicken embryonic SL-29 fibroblasts were transiently transfected with HA-tagged SDC4, fixed with 4% PFA, stained with mouse α-HA (left) and rabbit α-SDC4 (middle) followed by goat anti-mouse Alexa Fluor 488 and goat anti-rabbit Alexa Fluor 546 before fluorescence microscopy analyses. NucBlue was used to stain nuclei (blue). The inserts represent a higher magnification of the framed areas. Arrows indicate focal adhesions. Scalebar 50 µm. (B) SL-29 cells were transiently transfected with SDC4 (n = 6), or HA-SDC4 (n = 3) plasmids. Lipofectamine only was used as control (Ctrl). Bars show the relative mRNA gene expression in transfected cells compared with the mean average of Ctrl cells (n = 6). The bars are presented as ± SEM. Asterisks denote significant differences between Ctrl and transfected cells, statistics assessed using unpaired t-test with Welch correction (ns > 0.05; *p ≤ 0.05; **p ≤ 0.01). (C) A representative western blot showing the level of SDC4 after transient transfection of the chicken fibroblasts. Cells treated as in B were subjected to western blotting using antibodies against HA tag adding a molecular size increase around nine amino acid (top) or only the cytoplasmic part of SDC4 (middle panel). The lowermost panel shows total protein. (D) Quantification of the levels of the 15 and 20 kDa SDC4 fragments, normalized to total protein level. The bars represent n = 8 in one technical replicate for SDC4 and Ctrl, and n = 4 in one technical replicate for HA-SDC4. The bars are presented as ± SEM. Asterisks denote significant differences between Ctrl and transfected cells, statistics assessed using unpaired Mann-Whitney non-parametric t-test (ns > 0.05; **p ≤ 0.01; ***p ≤ 0.001).

Article Snippet: SDC4 , Gg03370419_m1.

Techniques: Over Expression, Transfection, Staining, Fluorescence, Microscopy, Control, Gene Expression, Western Blot, MANN-WHITNEY

Journal: Frontiers in Physiology

Article Title: Dual roles of syndecan-4 in regulating chicken fibrosis in vitro

doi: 10.3389/fphys.2026.1782914

Figure Lengend Snippet: Effect of SDC4 overexpression on fibrosis markers in chicken fibroblasts SL-29. Protein expression of (A) pro-collagen I beta chain (250 kDa) and alpha chain (100 kDa), (B) pro-collagen III (250 kDa) and collagen I (100 kDa), (C) MMP-2 and (D) MMP-9 in SDC4 transfected SL-29 cells. GAPDH was used as a reference protein. The bars represent n = 3 in one technical replicate. Gene expression (n = 6 in technical triplicates) of (E) COL1A1, COL3A1, MMP2, MMP9, (F) TGFβ and IL1β in SDC4 transfected SL-29 cells. The bars are presented as ± SEM. Asterisks denote significant differences between Ctrl (lipofectamine only) and transfected cells, statistics assessed using unpaired t-test with Welch correction and Mann-Whitney U (ns > 0.05; *p ≤ 0.05; **p ≤ 0.001).

Article Snippet: SDC4 , Gg03370419_m1.

Techniques: Over Expression, Expressing, Transfection, Gene Expression, MANN-WHITNEY

Journal: Frontiers in Physiology

Article Title: Dual roles of syndecan-4 in regulating chicken fibrosis in vitro

doi: 10.3389/fphys.2026.1782914

Figure Lengend Snippet: Effect of SDC4 overexpression on signaling pathways chicken fibroblasts. Protein expression analysis following SDC4 overexpression in SL-29 chicken fibroblasts. Detected proteins include (A) phosphorylated p38 at Thr186/Tyr182 and total p38, (B) phosphorylated Akt at Ser473 (pSer473- Akt) and total Akt, (C) phosphorylated ribosomal protein S6 at Ser240/244 (pSer240/244-S6) and total ribosomal protein S6, and (D) non-phosphorylated (active) β-catenin at Ser33/37 and Thr41 and total β-catenin. The ratio of phosphorylated to total or active to total protein was calculated for each protein, illustrating the impact of SDC4 overexpression. As a control (ctrl) was used treatment with lipofectamine only. GAPDH was used as a reference protein. The bars are presented as ± SEM from n = 3-6 in one technical replicate. Asterisks denote significant differences between Ctrl and transfected cells, statistics assessed using either unpaired t-test with Welch correction or Mann-Whitney U test (ns > 0.05; *p ≤ 0.05; **p ≤ 0.01).

Article Snippet: SDC4 , Gg03370419_m1.

Techniques: Over Expression, Protein-Protein interactions, Expressing, Control, Transfection, MANN-WHITNEY

Journal: Frontiers in Physiology

Article Title: Dual roles of syndecan-4 in regulating chicken fibrosis in vitro

doi: 10.3389/fphys.2026.1782914

Figure Lengend Snippet: Effect of blocking peptides on SDC4 shedding. (A) Alignment of human and chicken SDC4 protein sequences, highlighting amino acid percentage identity (dark blue >80%, lighter shades of blue are >60% and >40%, white <40%). Five overlapping blocking peptides (BP1-5) representing the SDC4 ectodomain were designed and are marked by different colors below the chicken SDC4 sequence. Red arrows indicate MMP cleavage sites in human SDC4 . Alignment created by Jalview 2.11.4.0. (B) Effect of BP1-5 on SDC4 shedding in SDC4-overexpressing chicken fibroblasts (N = 4 in one technical replicate). Total protein is used as a loading control. The bars on the right are presented as ± SEM. SDC4-overexpressing fibroblasts treated with the solvent of BP serve as the control (Ctrl, stippled line). Asterisks denote significant differences between Ctrl and transfected cells, statistics assessed using either unpaired t-test with Welch correction or Mann-Whitney U test (*p ≤ 0.05; **p ≤ 0.01).

Article Snippet: SDC4 , Gg03370419_m1.

Techniques: Blocking Assay, Sequencing, Control, Solvent, Transfection, MANN-WHITNEY

Journal: Frontiers in Physiology

Article Title: Dual roles of syndecan-4 in regulating chicken fibrosis in vitro

doi: 10.3389/fphys.2026.1782914

Figure Lengend Snippet: Effect of blocking peptides on the gene expression of the various SDCs. Effect of BP1-5 on the levels of SDC1-4 expression in SDC4 transfected chicken fibroblasts SL-29 showing no significant differences compared to control (Ctrl). All results were compared to the appropriate solvent for each blocking peptide. Gene expression was measured in three biological replicates, each performed in technical triplicates, and statistical significance was calculated using one-way ANOVA with Welch and Brown-Forsythe corrections. The bars are presented as ± SEM.

Article Snippet: SDC4 , Gg03370419_m1.

Techniques: Blocking Assay, Gene Expression, Expressing, Transfection, Control, Solvent

Journal: Frontiers in Physiology

Article Title: Dual roles of syndecan-4 in regulating chicken fibrosis in vitro

doi: 10.3389/fphys.2026.1782914

Figure Lengend Snippet: Effect of blocking peptides on the gene expression of fibrosis markers. Effect of BP1-5 on TGFB1, COL1A1, COL3A1, MMP9, MMP2, IL1B, ACTA2, DCN gene expression levels in SDC4 transfected chicken fibroblasts SL-29. All results were compared to the appropriate solvent for each blocking peptide. Gene expression was measured in three biological replicates, each performed in technical triplicates, and statistical significance was calculated using one-way ANOVA with Welch and Brown-Forsythe corrections. The bars are presented as ± SEM.

Article Snippet: SDC4 , Gg03370419_m1.

Techniques: Blocking Assay, Gene Expression, Transfection, Solvent

Journal: Frontiers in Physiology

Article Title: Dual roles of syndecan-4 in regulating chicken fibrosis in vitro

doi: 10.3389/fphys.2026.1782914

Figure Lengend Snippet: Effect of TGF-β1 on different gene and protein expression levels. Chicken fibroblasts were treated with TGF-β1 for 24 h. The gene expression levels were assessed for (A) syndecans (SDC1-4) and various fibrotic markers, including (B) COL1A1, (C) COL3A1, (D) MMP2, and (E) MMP9. Additionally, cytokines such as (F) TGFB1 and (G) IL1B were measured, and the expression of (H) ACTA2, a myofibroblast marker, and (I) DCN, a small leucine-rich proteoglycan. Results are expressed as fold change relative to the control (Ctrl; SL-29 cells treated with the solvent of TGF-β1). Gene expression (n = 7, in technical triplicates) was calculated by an unpaired t-test with Welch’s correction. ns > 0.05; *p ≤ 0.05; **p ≤ 0.01. The bars are presented as ± SEM. (J) Immunoblotting analysis of the 20 kDa SDC4 fragment following 24-h treatment with TGF-β1, compared to control (Ctrl) conditions. Quantification (N = 3 in one technical replicate) is presented as the mean, normalized to total protein, and expressed as a percentage relative to the control.

Article Snippet: SDC4 , Gg03370419_m1.

Techniques: Expressing, Gene Expression, Marker, Control, Solvent, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: AGO promotes lysosomal degradation of SDC4-CTF. A , Western blot of SDC1/4 CTF and full-length in SW480 cells treated with AGO (indicated doses, 24 h; n = 3). B , Western blot in SW480 cells treated with 20 μM AGO for 0 to 24 h. C and D , immunofluorescence detection and quantification of SDC4 in SW480 cells treated with 20 μM AGO (24 h; n = 6). E , surface SDC4 measured by FACS using FITC-anti-SDC4. F , Western blot of SDC1/4 CTF and full-length in HCT116 cells treated with AGO (indicated doses, 24 h). G , Western blot of SDC4-CTF in multiple colorectal cancer cells treated with 20 μM AGO (24 h). H , Western blot of SDC1-CTF and SDC4-CTF in SW480 cells treated with cycloheximide (50 μg/ml) along with/without 20 μM AGO for indicated times. I , RT-qPCR of indicated genes in SW480 cells treated with 20 μM AGO (24 h; n = 4). J , Western blot of SDC1-CTF and SDC4-CTF in SW480 cells treated with 20 μM AGO, 2.5 μM GM6001, 10 μM TMI-1, or combinations (24 h). K , Western blot in SW480 cells treated with 20 μM AGO, 5 μM BAY11 to 7082, 10 nM PMA, 1 μg/ml LPS, or 20 μg/ml TNFα (24 h). L and M , live-cell imaging and analysis of SDC4-GFP intensity in 293T cells co-expressing DsRed-Rab5A and SDC4-GFP, with 20 μM AGO treatment (n = 4). O , Western blot of lysosome markers and SDC1/4-CTF in SW480 cells treated with 20 μM AGO, 250 nM BAF1, or combination (24 h). Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗ p < 0.05, ∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, ns). AGO, andrographolide; SDC4, Syndecan4; CTF, C-terminal transmembrane fragment; FACS, fluorescence-activated cell sorting; BAF1, Bafilomycin A1.

Article Snippet: To measure surface populations of SDC4, resuspended cells (>10,000) were incubated with FITC-conjugated anti-SDC4 or its isotype-matched control antibody (sc-12766, Santa Cruz) for 15 min on ice and processed for flow cytometry analysis following the manufacturer’s instructions.

Techniques: Western Blot, Immunofluorescence, Quantitative RT-PCR, Live Cell Imaging, Expressing, Two Tailed Test, Fluorescence, FACS

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: AGO promotes lysosomal degradation of SDC4-CTF. A , Western blot of SDC1/4 CTF and full-length in SW480 cells treated with AGO (indicated doses, 24 h; n = 3). B , Western blot in SW480 cells treated with 20 μM AGO for 0 to 24 h. C and D , immunofluorescence detection and quantification of SDC4 in SW480 cells treated with 20 μM AGO (24 h; n = 6). E , surface SDC4 measured by FACS using FITC-anti-SDC4. F , Western blot of SDC1/4 CTF and full-length in HCT116 cells treated with AGO (indicated doses, 24 h). G , Western blot of SDC4-CTF in multiple colorectal cancer cells treated with 20 μM AGO (24 h). H , Western blot of SDC1-CTF and SDC4-CTF in SW480 cells treated with cycloheximide (50 μg/ml) along with/without 20 μM AGO for indicated times. I , RT-qPCR of indicated genes in SW480 cells treated with 20 μM AGO (24 h; n = 4). J , Western blot of SDC1-CTF and SDC4-CTF in SW480 cells treated with 20 μM AGO, 2.5 μM GM6001, 10 μM TMI-1, or combinations (24 h). K , Western blot in SW480 cells treated with 20 μM AGO, 5 μM BAY11 to 7082, 10 nM PMA, 1 μg/ml LPS, or 20 μg/ml TNFα (24 h). L and M , live-cell imaging and analysis of SDC4-GFP intensity in 293T cells co-expressing DsRed-Rab5A and SDC4-GFP, with 20 μM AGO treatment (n = 4). O , Western blot of lysosome markers and SDC1/4-CTF in SW480 cells treated with 20 μM AGO, 250 nM BAF1, or combination (24 h). Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗ p < 0.05, ∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, ns). AGO, andrographolide; SDC4, Syndecan4; CTF, C-terminal transmembrane fragment; FACS, fluorescence-activated cell sorting; BAF1, Bafilomycin A1.

Article Snippet: Proximity ligation assay was performed according to the protocol provided by Sigma-Aldrich (DUO092101), and the primary antibodies and dilutions are anti-SDC4 (sc12766, 1:100), anti-Syntenin (22399-1-AP, 1:400, Proteintech).

Techniques: Western Blot, Immunofluorescence, Quantitative RT-PCR, Live Cell Imaging, Expressing, Two Tailed Test, Fluorescence, FACS

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: MG132 inhibits the degradation of SDC4-CTF. A and B , Western blot of HCT116 cells treated with 0.5, 1, 2.5, or 5 μM MG132 for 12 h, or with 2.5 μM MG132 for 0 to 12 h. C , Western blot of full-length SDC1 and SDC4 in HCT116 cells treated with MG132 (0.5–5 μM, 12 h). D and E , immunofluorescence detection and quantification of HCT116 cells transfected with SDC1-GFP or SDC4-GFP, with/without 5 μM MG132 (4 h; n = 6). F , Western blot of multiple colorectal cancer cell lines treated with 10 μM MG132 (12 h). G , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with 0.1, 0.5, or 1 μM proteasome inhibitors (Carfilzomib, Ixazomib, Bortezomib) for 24 h. H , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with PD150606 (2, 5, 10 μM) for 12 h. I , quantification of DQ-BSA fluorescence with MG132 or Earle’s balanced salt solution (EBSS) treatment (n = 4). J , quantification of lysosomal activity using LysoTracker with MG132 or EBSS treatment (n = 4). K , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells with siRNA knockdown of proteasome subunits PSMD14, PSMD2, USP14, PSMB5, PSMA6. Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001). SDC4, syndecan4; CTF, C-terminal transmembrane (TM) fragment; EBSS, Earle’s balanced salt solution.

Article Snippet: SDC4 rabbit polyclonal antibody (NB110–41551) was purchased from Novus for SDC4-CTF detection.

Techniques: Western Blot, Immunofluorescence, Transfection, Fluorescence, Activity Assay, Knockdown, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: SDC4-CTF is further cleaved by γ-secretase to release the cytoplasmic fragment. A , confocal microscopy of HCT116 cells transfected with SDC4-GFP or SDC4-CTF-GFP. B , schematic of SDC4-GFP deletion mutants, SNP for Signal peptide, ED for ectodomain, TM for TM motif, C1 and C2 for the constant regions, and V for variable region. C , Western blot of HCT116 cells transfected with SDC1-GFP, SDC4-GFP, or SDC4-CTF-GFP, with/without 5 μM MG132 (4 h). D , sequence alignment of Syndecan family TM C-terminal fragments. The alignment was performed using Clustal W, and analysis was done with ESPript 3.0. E , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells with presenilin-1 knockdown (siRNA, 72 h). F , Western blot of HCT116 and SW480 cells treated with 10 μM DAPT, 10 μM MG132, or 2.5 μM GM6001 (12 h). G , schematic of SDC4-CTF degradation via lysosome or proteasome pathways. Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗∗ p < 0.001, ∗∗∗ p < 0.0005, ∗∗∗∗ p < 0.0001). SDC4, Syndecan4; CTF, C-terminal transmembrane fragment.

Article Snippet: SDC4 rabbit polyclonal antibody (NB110–41551) was purchased from Novus for SDC4-CTF detection.

Techniques: Confocal Microscopy, Transfection, Western Blot, Sequencing, Knockdown, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: Syntenin stabilizes SDC4-CTF against endocytic-lysosomal degradation. A , Western blot of SDC1-CTF and SDC4-CTF in HCT116 cells treated with 25 μM LY294002, 250 nM Rapamycin, or 250 nM Bafilomycin A1. B , Western blot of HCT116 cells starved in EBSS for indicated durations. C and D , Western blot and quantification of SDC1-CTF and SDC4-CTF in HCT116 cells overexpressing Flag-tagged Syntenin (n = 3). E , schematic and sequences of SDC4 WT and SDC4 ΔC2 , with confocal images of indicated plasmids in 293T cells. F , quantification of fluorescence co-localization from ( E ) (n = 6). G and H , Western blot and quantification of SDC4-CTF and Syntenin in 293T cells co-transfected with SDC4 and GFP-tagged Syntenin (n = 3). I , Western blot of SDC1-CTF, SDC4-CTF, and Syntenin in multiple cancer cell lines. J , correlation analysis between SDC4-CTF and Syntenin expression from ( I ). Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗∗ p < 0.001, ∗∗∗ p < 0.0005, ns). SDC4, Syndecan4; CTF, C-terminal transmembrane fragment; EBSS, Earle’s balanced salt solution.

Article Snippet: SDC4 rabbit polyclonal antibody (NB110–41551) was purchased from Novus for SDC4-CTF detection.

Techniques: Western Blot, Fluorescence, Transfection, Expressing, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: AGO binds to SDC4 and disrupts SDC4-Syntenin interaction. A , chemical structure of AGO. B , microscale thermophoresis (MST) assays of AGO with GFP-SDC4 or GFP control in 293T lysate (n = 3). C , MST assays of AGO with recombinant glutathione S-transferase (GST)-SDC4 (n = 3). D and E , MST assays of AGO with GFP-tagged SDC4 or SDC1 and their TM deletion mutants (n = 3 each). F and G , Western blot and quantification of AGO drug affinity responsive target stability experiments (n = 3). H and I , immunoprecipitation (IP) and quantification of SDC4 binding to Syntenin in SW480 cells treated with 20 μM AGO (24 h; n = 3). J , IP of Flag-Syntenin and GFP-SDC4 in 293T cells treated with 20 μM AGO (24 h). K , GST pull-down of Syntenin from 293T lysate with/without 20 μM AGO, using GST or GST-SDC4. L and M , poximity ligation assays and quantification of SDC4-Syntenin interaction in SW480 cells treated with 20 μM AGO (24 h; n = 8). O and P , confocal images and quantification of SDC4/GFP-Syntenin co-localization in 293T cells treated with 20 μM AGO (24 h; n = 8). Q , IP of SDC4 and detection of GFP-Syntenin/endogenous Syntenin in 293T cells treated with 20 μM AGO (24 h). Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗∗ p < 0.001, ∗∗∗ p < 0.0005, ns). AGO, Andrographolide; GST, glutathione S-transferase; SDC4, Syndecan4; MST, microscale thermophoresis; TM, transmembrane; IP, immunoprecipitation.

Article Snippet: SDC4 rabbit polyclonal antibody (NB110–41551) was purchased from Novus for SDC4-CTF detection.

Techniques: Microscale Thermophoresis, Control, Recombinant, Western Blot, Immunoprecipitation, Binding Assay, Ligation, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Andrographolide targets syndecan4 to impair its interaction with syntenin and inhibits the biogenesis of small extracellular vesicles

doi: 10.1016/j.jbc.2026.111182

Figure Lengend Snippet: AGO promotes lysosomal degradation of SDC4-CTF. A , Western blot of SDC1/4 CTF and full-length in SW480 cells treated with AGO (indicated doses, 24 h; n = 3). B , Western blot in SW480 cells treated with 20 μM AGO for 0 to 24 h. C and D , immunofluorescence detection and quantification of SDC4 in SW480 cells treated with 20 μM AGO (24 h; n = 6). E , surface SDC4 measured by FACS using FITC-anti-SDC4. F , Western blot of SDC1/4 CTF and full-length in HCT116 cells treated with AGO (indicated doses, 24 h). G , Western blot of SDC4-CTF in multiple colorectal cancer cells treated with 20 μM AGO (24 h). H , Western blot of SDC1-CTF and SDC4-CTF in SW480 cells treated with cycloheximide (50 μg/ml) along with/without 20 μM AGO for indicated times. I , RT-qPCR of indicated genes in SW480 cells treated with 20 μM AGO (24 h; n = 4). J , Western blot of SDC1-CTF and SDC4-CTF in SW480 cells treated with 20 μM AGO, 2.5 μM GM6001, 10 μM TMI-1, or combinations (24 h). K , Western blot in SW480 cells treated with 20 μM AGO, 5 μM BAY11 to 7082, 10 nM PMA, 1 μg/ml LPS, or 20 μg/ml TNFα (24 h). L and M , live-cell imaging and analysis of SDC4-GFP intensity in 293T cells co-expressing DsRed-Rab5A and SDC4-GFP, with 20 μM AGO treatment (n = 4). O , Western blot of lysosome markers and SDC1/4-CTF in SW480 cells treated with 20 μM AGO, 250 nM BAF1, or combination (24 h). Data: mean ± SD (≥3 experiments). Statistics: unpaired two-tailed t test (∗ p < 0.05, ∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, ns). AGO, andrographolide; SDC4, Syndecan4; CTF, C-terminal transmembrane fragment; FACS, fluorescence-activated cell sorting; BAF1, Bafilomycin A1.

Article Snippet: SDC4 rabbit polyclonal antibody (NB110–41551) was purchased from Novus for SDC4-CTF detection.

Techniques: Western Blot, Immunofluorescence, Quantitative RT-PCR, Live Cell Imaging, Expressing, Two Tailed Test, Fluorescence, FACS

Journal: Cartilage

Article Title: Effect of Synovial Cell Fractionation on Tenascin-C Expression in Chondrocytes Under Coculture

doi: 10.1177/19476035261421469

Figure Lengend Snippet: Gene expression in the CD68 positive group. (A) Chondrocyte mRNA expression. (B) mRNA expression in synovial cells. Columns and bars indicate means and standard deviations. Coculture upregulated TNC and MMP-3 expression in chondrocytes and SDC4 expression in synovial cells. AC = chondrocytes; AC_S = chondrocytes cocultured with synovial cells; S = synovial cells; S_AC = synovial cells cocultured with chondrocytes; MMP = matrix metalloproteinase; TNC = tenascin-C; SDC4 = syndecan-4; FN = fibronectin; TIMP3 = tissue inhibitor of metalloproteinase 3; TGF = transforming growth factor; FGF = fibroblast growth factor; ADAMTS5 = a disintegrin and metalloproteinase with thrombospondin motifs 5. * P < 0.05.

Article Snippet: TaqMan gene expression assay primer-probe pairs were obtained for the detection of TNC (assay no. Hs01115665-m1), SDC4 (assay no. Hs00161617-m1), fibronectin (assay no. Hs01549976-m1), TGF-β (assay no. Hs00998133-m1), TIMP 3 (assay no. Hs00165949-m1), bFGF (assay no. Hs00256645-m1), ADAMTS 5 (assay no. Hs00199841-m1), MMP-3 (assay no. Hs00968305-m1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (assay no. Hs99999905-m1), IL-1β (assay no. Hs01555410-m1), and TNFα (assay no. Hs01113624-g1).

Techniques: Gene Expression, Expressing

Journal: Cartilage

Article Title: Effect of Synovial Cell Fractionation on Tenascin-C Expression in Chondrocytes Under Coculture

doi: 10.1177/19476035261421469

Figure Lengend Snippet: Gene expression in the CD68 negative group. (A) Chondrocyte mRNA expression. (B) mRNA expression in synovial cells. Columns and bars indicate means and standard deviations. Coculture did not significantly alter the mRNA expression of each factor in chondrocytes or synovial cells. AC = chondrocytes; AC_S = chondrocytes cocultured with synovial cells; S = synovial cells; S_AC = synovial cells cocultured with chondrocytes; MMP = matrix metalloproteinase; TNC = tenascin-C; SDC4 = syndecan-4; FN = fibronectin; TIMP3 = tissue inhibitor of metalloproteinase 3; TGF = transforming growth factor; FGF = fibroblast growth factor; ADAMTS5 = a disintegrin and metalloproteinase with thrombospondin motifs 5.

Article Snippet: TaqMan gene expression assay primer-probe pairs were obtained for the detection of TNC (assay no. Hs01115665-m1), SDC4 (assay no. Hs00161617-m1), fibronectin (assay no. Hs01549976-m1), TGF-β (assay no. Hs00998133-m1), TIMP 3 (assay no. Hs00165949-m1), bFGF (assay no. Hs00256645-m1), ADAMTS 5 (assay no. Hs00199841-m1), MMP-3 (assay no. Hs00968305-m1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (assay no. Hs99999905-m1), IL-1β (assay no. Hs01555410-m1), and TNFα (assay no. Hs01113624-g1).

Techniques: Gene Expression, Expressing

Journal: Pathophysiology

Article Title: Metabolic Syndrome-Driven Changes in Cardiac Lymphatic Endothelium: mRNA Expression and Emerging Questions

doi: 10.3390/pathophysiology33010004

Figure Lengend Snippet: RT-PCR analysis. Selected mRNA expression in sorted LECs from db/db and control mice hearts. Some mRNA levels significantly increase in db/db mice compared to controls. Statistically significant differences ( p -value < 0.05) are marked (*). Abbreviations: Acta2—Actin 2/smooth muscle actin; Bax—BCL2 Associated X; Bcl2—B-cell CLL/lymphoma 2; CCL21—Chemokine (C-C motif) ligand 21; CEACAM1—Carcinoembryonic antigen-related cell adhesion molecule 1; CPT1A—Carnitine Palmitoyltransferase 1A; CX3CL1—chemokine (C-X3-C motif) ligand 1; eNOS—endothelial nitric oxide synthase;Fbn1—fibrillin-1; FOXO1—Forkhead Box O1; GATA2—GATA-binding factor 2; ICAM-1—intercellular adhesion molecule 1; KLF2—Krüppel-like Factor 2; MMP2—Metalloproteinase 2; PDPN—podoplanin; PTEN—phosphatase and tensin homolog deleted on chromosome ten; Reln—reelin; Sdc4—syndecan 4; Snail1—Snail family transcriptional repressor-1; Snail2—Snail family transcriptional repressor-2; VCAM-1—vascular cell adhesion molecule 1; VE-cadherin—Vascular Endothelial Cadherin.

Article Snippet: To detect mRNA levels of selected genes, the following TaqMan Gene Expression Assays were used: CD36: Mm00432403_m1; Pten: Mm00477208_m1; Foxo1: Mm00490671_m1; Klf2: Mm00500486_g1; Bax: Mm00432051_m1; Bcl2: Mm00477631_m1; Cpt1a: Mm01231183_m1; Ccl21a: Mm03646971_gH; Cx3cl1: Mm00436454_m1; Icam1: Mm00516023_m1; Vcam1: Mm01320970_m1; Emilin1: Mm00467244_m1; Mmp2: Mm00439498_m1; Reln: Mm00465200_m1; Snai1: Mm00441533_g1; Snai2: Mm00441531_m1; Acta2: Mm00725412_s1; Cdh5: Mm00546194_s1; Gata2: Mm00492301_m1; Cldn5: Mm00727012_s1; Ceacam1: Mm04204476_m1; Pdpn: Mm01348912_g1; Fbn1: Mm00435217_m1; Nos3: Mm00435217_m1; Sdc4: Mm00488527_m1.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Control, Binding Assay

Journal: Pathophysiology

Article Title: Metabolic Syndrome-Driven Changes in Cardiac Lymphatic Endothelium: mRNA Expression and Emerging Questions

doi: 10.3390/pathophysiology33010004

Figure Lengend Snippet: Myocardial perivascular LVs express Sdc4 (red) in db/db mice ( a ). ( b ) shows relative quantification of mRNA level for Sdc4. Scale bar—50 µm. Abbreviations: Sdc4—syndecan 4. Statistically significant differences ( p -value < 0.05) are marked (*).

Article Snippet: To detect mRNA levels of selected genes, the following TaqMan Gene Expression Assays were used: CD36: Mm00432403_m1; Pten: Mm00477208_m1; Foxo1: Mm00490671_m1; Klf2: Mm00500486_g1; Bax: Mm00432051_m1; Bcl2: Mm00477631_m1; Cpt1a: Mm01231183_m1; Ccl21a: Mm03646971_gH; Cx3cl1: Mm00436454_m1; Icam1: Mm00516023_m1; Vcam1: Mm01320970_m1; Emilin1: Mm00467244_m1; Mmp2: Mm00439498_m1; Reln: Mm00465200_m1; Snai1: Mm00441533_g1; Snai2: Mm00441531_m1; Acta2: Mm00725412_s1; Cdh5: Mm00546194_s1; Gata2: Mm00492301_m1; Cldn5: Mm00727012_s1; Ceacam1: Mm04204476_m1; Pdpn: Mm01348912_g1; Fbn1: Mm00435217_m1; Nos3: Mm00435217_m1; Sdc4: Mm00488527_m1.

Techniques: Quantitative Proteomics