antibody sdc3  (R&D Systems)

Journal: Cells

Article Title: The Role of the Cell Surface Heparan Sulfate Proteoglycan Syndecan-3 in Breast Cancer Pathophysiology

doi: 10.3390/cells14201612

Figure Lengend Snippet: ( A ) Pan-cancer analysis of SDC3 expression across human malignancies. RNA-sequencing data from The Cancer Genome Atlas (TCGA) reveals median SDC3 expression levels across 31 cancer types, normalized as fragments per kilobase of exon per million mapped reads (FPKM). ( B ) SDC3 expression in breast cancer histological subtypes. Immunohistochemical staining of SDC3 in human breast tumor specimens presents as membranous and cytoplasmic staining that is primarily found in tumor cells, and less pronounced in the stroma. Scale bars are available at the original TCGA website.

Article Snippet: Cells were blocked for 1 h and then stained overnight at 4 °C with the primary antibody SDC3 (R&D Systems, Minneapolis, MN, USA; goat anti-Human Syndecan-3 Isoform 1 Antibody, 1:150, Cat. #: AF3539), and Vinculin (Sigma-Aldrich, Anti-Vinculin Antibody mouse monoclonal, 1:150, Cat. #: V9131).

Techniques: Expressing, RNA Sequencing, Immunohistochemical staining, Staining

Journal: Cells

Article Title: The Role of the Cell Surface Heparan Sulfate Proteoglycan Syndecan-3 in Breast Cancer Pathophysiology

doi: 10.3390/cells14201612

Figure Lengend Snippet: SDC3 is highly expressed in tumors and its expression correlates with better survival. ( A ) SDC3 expression varies across breast cancer progression stages. Gene chip analysis reveals significantly increased SDC3 mRNA levels in primary tumors and metastatic lesions, compared to normal breast tissue. ( B ) SDC3 expression is associated with the relapse-free survival ( RFS ) of breast cancer patients. Kaplan–Meier relapse-free survival curves are plotted based on the following: all breast cancer patients ( n = 4929), HER2 negative status ( n = 4047), St. Gallen luminal A status ( n = 2277), Grade 2 ( n = 1177), and neoadjuvant chemotherapy ( n = 402). Log-rank p values and hazard ratios (HRs; 95% confidence interval in parentheses) are shown.

Article Snippet: Cells were blocked for 1 h and then stained overnight at 4 °C with the primary antibody SDC3 (R&D Systems, Minneapolis, MN, USA; goat anti-Human Syndecan-3 Isoform 1 Antibody, 1:150, Cat. #: AF3539), and Vinculin (Sigma-Aldrich, Anti-Vinculin Antibody mouse monoclonal, 1:150, Cat. #: V9131).

Techniques: Expressing

Journal: Cells

Article Title: The Role of the Cell Surface Heparan Sulfate Proteoglycan Syndecan-3 in Breast Cancer Pathophysiology

doi: 10.3390/cells14201612

Figure Lengend Snippet: SDC3 is expressed differentially in several breast cancer cell lines and alters their metabolic activity and cell cycle. ( A ) Relative gene expression of SDC3 was quantified by qRT-PCR in 10 different breast cancer cell lines, representative of the luminal A (MCF-7), luminal B (BT474), HER2-positive (SKBR3, MDA-MB 453), triple-negative A (MDA-MB 468, HCC1806, BT20), and triple-negative B (MDA-MB-231, BT549, SUM149) subtype. Individual experiments were normalized against β-Actin and the relative expression was represented by 2-ΔCt. ( B ) SDC3 is downregulated in MDA-MB-231, and MCF-7 cells following SDC3 siRNA- and siPool-transfection. SDC3 knockdown was confirmed by qRT-PCR. ( C – E ) The downregulation of SDC3 was further assessed by Western blot ( C ), flow cytometry ( D ), and immunostaining for SDC3 (green fluorescence), vinculin (red), and nuclear staining with DAPI (blue) ( E ). The analysis revealed a reduced expression of SDC3 in KD cells ( D ), as well as its constitutive cellular localization. In control cells, SDC3 was predominantly localized in the cytoplasm, nucleus, and in focal adhesion-like structures. In contrast, KD cells exhibited a loss of focal adhesion-like and cytoplasmic staining of SDC3 ( E ). Inserts highlight magnified regions of individual cells, indicating SDC3 expression and vinculin in structures resembling focal adhesions of control cells and the absence of these structures in KD cells. p ≤ 0.01 was described as very significant (**), and p ≤ 0.001 as highly significant (***). The graphs depict mean values, with error bars indicating the standard error of the mean (SEM). Three independent experimental replicates were included ( n = 3); original magnification 400×.

Article Snippet: Cells were blocked for 1 h and then stained overnight at 4 °C with the primary antibody SDC3 (R&D Systems, Minneapolis, MN, USA; goat anti-Human Syndecan-3 Isoform 1 Antibody, 1:150, Cat. #: AF3539), and Vinculin (Sigma-Aldrich, Anti-Vinculin Antibody mouse monoclonal, 1:150, Cat. #: V9131).

Techniques: Activity Assay, Gene Expression, Quantitative RT-PCR, Expressing, Transfection, Knockdown, Western Blot, Flow Cytometry, Immunostaining, Fluorescence, Staining, Control

Journal: Cells

Article Title: The Role of the Cell Surface Heparan Sulfate Proteoglycan Syndecan-3 in Breast Cancer Pathophysiology

doi: 10.3390/cells14201612

Figure Lengend Snippet: SDC3 alters the metabolic activity and cell cycle of breast cancer cells. ( A ) SDC3 depletion affects the metabolic activity of MDA-MB-231 and MCF7 breast cancer cells. Breast cancer cells were subjected to the metabolic MTT assay following transfections with the siRNA and siPool. ( B ) SDC3-knockdown promotes cell cycle progression in MDA-MB-231 cells. Cell cycle phase composition was measured employing DAPI staining and flow cytometry after SDC3 depletion. p ≤ 0.05 was considered statistically significant (*), while p ≤ 0.01 was described as very significant (**). The graphs depict mean values, with error bars indicating the standard error of the mean (SEM). Three independent experimental replicates were included ( n = 3).

Article Snippet: Cells were blocked for 1 h and then stained overnight at 4 °C with the primary antibody SDC3 (R&D Systems, Minneapolis, MN, USA; goat anti-Human Syndecan-3 Isoform 1 Antibody, 1:150, Cat. #: AF3539), and Vinculin (Sigma-Aldrich, Anti-Vinculin Antibody mouse monoclonal, 1:150, Cat. #: V9131).

Techniques: Activity Assay, MTT Assay, Transfection, Knockdown, Staining, Flow Cytometry

Journal: Cells

Article Title: The Role of the Cell Surface Heparan Sulfate Proteoglycan Syndecan-3 in Breast Cancer Pathophysiology

doi: 10.3390/cells14201612

Figure Lengend Snippet: SDC3 depletion affects the growth of 3D spheroids in MDA-MB 231 and MCF-7 cells. Representative pictures of the hanging drop cultures are presented in the left panels and quantitative analysis in the right panels. Spheroid area was quantified using NIH Image J software and expressed as arbitrary units (AU). ( A ) MDA-MB-231 cells were transfected with the negative control, SDC3 siRNA, and SDC3 siPool, and subjected to the hanging drop assay. ( B ) MCF7 cells were transfected with the negative control, SDC3 siRNA, and SDC3 siPool, and subjected to the hanging drop assay. p ≤ 0.05 was considered statistically significant (*), and p ≤ 0.001 as highly significant (***). The graphs depict mean values, with error bars indicating the standard error of the mean (SEM). Three independent experimental replicates were included ( n = 3).

Article Snippet: Cells were blocked for 1 h and then stained overnight at 4 °C with the primary antibody SDC3 (R&D Systems, Minneapolis, MN, USA; goat anti-Human Syndecan-3 Isoform 1 Antibody, 1:150, Cat. #: AF3539), and Vinculin (Sigma-Aldrich, Anti-Vinculin Antibody mouse monoclonal, 1:150, Cat. #: V9131).

Techniques: Software, Transfection, Negative Control

Journal: Cells

Article Title: The Role of the Cell Surface Heparan Sulfate Proteoglycan Syndecan-3 in Breast Cancer Pathophysiology

doi: 10.3390/cells14201612

Figure Lengend Snippet: ( A ) The migration of MDA-MB-231 cells was significantly decreased in transwell migration assays (10× magnification) and the ( B ) wound area was significantly increased in ibidi chamberwound closure assays following SDC3 knockdown, in comparison to the negative control. MCF-7 cells displayed little changes in migratory behavior or in wound area upon SDC3 depletion. Red line indicates maximum wound closure. 10× magnification.( C , D ) SDC3 regulates the expression of different potential effector genes. SDC3 depletion was achieved by siRNA- and siPool-mediated knockdown of SDC3 in MDA-MB-231 ( C ), and MCF-7 ( D ) cells. The expression of target genes associated with notch-signaling ( HES1 ), hedgehog-signaling ( Gli2 ), epithelial-to-mesenchymal-transition signaling ( Twist , CDH1 ), matrix-metalloproteinase signaling ( MMP1 , MMP2 , MMP9 ), and vascular endothelial growth factor signaling ( VEGF-A ) was visibly altered ( E ) The effect of SDC3 depletion on the potential activation of Src proto-oncogene tyrosine-protein kinase (pSRC), as well as expression of its total form ( SRC ) was examined using Western blot analysis. Upper panels = representative Western blots. Lower panels = densitometric quantification of three independent experiments. p ≤ 0.05 was considered statistically significant (*), while p ≤ 0.01 was described as very significant (**). The graphs depict mean values, with error bars indicating the standard error of the mean (SEM). Three independent experimental replicates were included ( n = 3).

Article Snippet: Cells were blocked for 1 h and then stained overnight at 4 °C with the primary antibody SDC3 (R&D Systems, Minneapolis, MN, USA; goat anti-Human Syndecan-3 Isoform 1 Antibody, 1:150, Cat. #: AF3539), and Vinculin (Sigma-Aldrich, Anti-Vinculin Antibody mouse monoclonal, 1:150, Cat. #: V9131).

Techniques: Migration, Knockdown, Comparison, Negative Control, Expressing, Activation Assay, Western Blot

Journal: Cells

Article Title: The Role of the Cell Surface Heparan Sulfate Proteoglycan Syndecan-3 in Breast Cancer Pathophysiology

doi: 10.3390/cells14201612

Figure Lengend Snippet: Functional analysis of SDC3 using STRING database. ( A ) STRING database output revealed direct connections between SDC3, SRC , FGF2 , and MMP7. Further indirect associations were established between SDC3 and FLT1 , KDR , VEGFB , VEGFC , CDH1 , MMP1 , MMP2 , MMP9 , and MMP14 , as well TIMP2 and TIMP4 . ( B ) In the gene ontology (GO) analysis, the 10 most significantly ( p < 0.05) enriched GO terms were included in the biological process (blue), molecular function (green), and cellular component (orange) categories. X-axis: −log10 (false discovery value) ( C ) KEGG-pathway analysis depicts several interconnections between SDC3 and relevant signaling pathways in cancer.

Article Snippet: Cells were blocked for 1 h and then stained overnight at 4 °C with the primary antibody SDC3 (R&D Systems, Minneapolis, MN, USA; goat anti-Human Syndecan-3 Isoform 1 Antibody, 1:150, Cat. #: AF3539), and Vinculin (Sigma-Aldrich, Anti-Vinculin Antibody mouse monoclonal, 1:150, Cat. #: V9131).

Techniques: Functional Assay, Protein-Protein interactions

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Syndecan-3 positively regulates the pro-inflammatory function of macrophages

doi: 10.1007/s00018-025-05649-1

Figure Lengend Snippet: SDC3, but not the other syndecans, is upregulated in response to IFNγ in human macrophages. ( a ) SDC1 , SDC2 , SDC3 and SDC4 mRNA levels in control and IFNγ-treated THP-1 macrophages, as assessed by RT-qPCR ( n = 4–6). ( b ) Representative flow histograms (upper panel) and pooled data (lower panel) showing the protein expression of SDC1, SDC2, SDC3 and SDC4 in control and IFNγ-treated THP-1 macrophages ( n = 4–6). ( c ) Representative confocal images ( n = 3) of control, IFNγ- or IL-4/IL-13-treated THP-1 macrophages immunostained for DAPI and SDC3 (scale bar 5 μm). ( d ) SDC3 mRNA levels in control, IFNγ- and IL-4/IL-13-treated THP-1 macrophages, as assessed by RT-qPCR ( n = 3). Means ± SEM. Statistically significant difference from controls or between indicated groups is shown by ns: not significant, ** p < 0.01, *** p < 0.001 and **** p < 0.0001

Article Snippet: SDC3 primary antibody (R&D Systems Cat. #: AF3539), was incubated overnight at 4 °C in blocking buffer.

Techniques: Control, Quantitative RT-PCR, Expressing

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Syndecan-3 positively regulates the pro-inflammatory function of macrophages

doi: 10.1007/s00018-025-05649-1

Figure Lengend Snippet: SDC3 deficient macrophages exhibit aberrant proliferation, adhesion and expression of cell surface markers. ( a ) Representative immunoblots ( n = 3) of WT or SDC3 KO THP-1 macrophages stimulated with either 100 ng/ml of IFNγ or 20 ng/ml of IL-4/IL-13 cocktail. ( b ) SDC3 mRNA levels in control, IFNγ- and IL-4/IL-13-treated WT or SDC3 KO THP-1 macrophages, as assessed by RT-qPCR ( n = 3) ( c ) Representative brightfield images of WT or SDC3 KO THP-1 macrophages (scale bar 20 μm, n = 3). ( d ) Proliferation of WT or SDC3 KO THP-1 cells up to 5 days ( n = 3). ( e ) Adhesion activity of WT or SDC3 KO THP-1 macrophages, as quantified by Crystal Violet ( n = 2–3). ( f ) Flow cytometry data showing the expression of CD40, CD86 and CD163 in WT or SDC3 KO THP-1 macrophages stimulated with 100 ng/ml of IFNγ ( n = 7). Means ± SEM. Statistically significant difference from controls or between indicated groups is shown by ns: not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001

Article Snippet: SDC3 primary antibody (R&D Systems Cat. #: AF3539), was incubated overnight at 4 °C in blocking buffer.

Techniques: Expressing, Western Blot, Control, Quantitative RT-PCR, Activity Assay, Flow Cytometry

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Syndecan-3 positively regulates the pro-inflammatory function of macrophages

doi: 10.1007/s00018-025-05649-1

Figure Lengend Snippet: SDC3 -defective macrophages show distinctive gene expression patterns. ( a ) Heatmap of differentially expressed genes in IFNγ-treated THP-1 WT and SDC3 KO macrophages ( n = 3, purple represents upregulation and green represents downregulation). ( b ) Differentially expressed pathways in IFNγ-treated THP-1 WT and SDC3 KO macrophages where blue represents pathways upregulated in SDC3 KO macrophages, and orange represents downregulated pathways in SDC3 KO macrophages ( n = 3). ( c ) TNF , IL10 , iNOS , PD-L1 , CD86 , and VEGFA mRNA levels in IFNγ-treated THP-1 WT and SDC3 KO macrophages, as assessed by RT-qPCR ( n = 3). Means ± SEM. Statistically significant difference from controls or between indicated groups is shown by * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001

Article Snippet: SDC3 primary antibody (R&D Systems Cat. #: AF3539), was incubated overnight at 4 °C in blocking buffer.

Techniques: Gene Expression, Quantitative RT-PCR

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Syndecan-3 positively regulates the pro-inflammatory function of macrophages

doi: 10.1007/s00018-025-05649-1

Figure Lengend Snippet: SDC3 promotes macrophage phagocytic capacity and inhibits tumour-spheroid formation. ( a ) Left: Representative images of IFNγ-stimulated WT or SDC3 KO THP-1 macrophages co-cultured with MDA-MB-231-GFP breast cancer cells. Images were taken with a Leica SP8 Lightning confocal microscope (scale bar 20.5 μm, n = 3). Right: Quantification of phagocytosis rate by flow cytometry normalized to WT, using the inhibitor of actin polymerization Cytochalasin D as control ( n = 3). ( b ) Left: Representative images of IFNγ-treated WT or SDC3 KO THP-1 macrophages forming spheroids with MDA-MB-231-GFP cells. Images were taken using Nikon Eclipse TD 100 microscope at the indicated times (scale bar 100 μm, white colour represents GFP + tumour cells). Right: Proliferation of MDA-MB-231-GFP + tumour cells in each condition normalized to MDA-MB-231 condition, calculated after acquiring single cell suspensions of spheroids by flow cytometry ( n = 3). Means ± SEM. Statistically significant difference from controls or between indicated groups is shown by * p < 0.05, ** p < 0.01 and **** p < 0.0001

Article Snippet: SDC3 primary antibody (R&D Systems Cat. #: AF3539), was incubated overnight at 4 °C in blocking buffer.

Techniques: Cell Culture, Microscopy, Flow Cytometry, Control

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Syndecan-3 positively regulates the pro-inflammatory function of macrophages

doi: 10.1007/s00018-025-05649-1

Figure Lengend Snippet: SDC3 plays a role in macrophage pro-inflammatory functions. ( a ) Left: Representative confocal microscopy images of WT or SDC3 KO THP-1 macrophages stimulated with 100 ng/ml of IFNγ and incubated with pHrodo™ Green S. aureus bioparticles. Images were taken using a Leica SP8 Lightning confocal microscope (scale bar 10 μm). Right: Flow cytometry quantification of S. aureus phagocytosis normalized to WT, using the inhibitor of actin polymerization Cytochalasin D as control ( n = 3). ( b ) Cytokine quantification in cell supernatants from WT or SDC3 KO THP-1 macrophages stimulated with 100 ng/ml of IFNγ, using the LEGENDplex™ HU Essential Immune Response Panel ( n = 5). ( c ) Quantification of T cell proliferation index using CellTrace CFSE staining ( n = 5) in CD8 + T cells co-cultured with IFNγ-treated WT or SDC3 KO THP-1 macrophages. ( d ) Quantification of T cell activation markers PD-1 and CD95 by flow cytometry in CD8 + T cells co-cultured with IFNγ-treated WT or SDC3 KO THP-1 macrophages ( n = 5). Means ± SEM. Statistically significant difference from controls or between indicated groups is shown by ns: not significant, * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001

Article Snippet: SDC3 primary antibody (R&D Systems Cat. #: AF3539), was incubated overnight at 4 °C in blocking buffer.

Techniques: Confocal Microscopy, Incubation, Microscopy, Flow Cytometry, Control, Staining, Cell Culture, Activation Assay

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Syndecan-3 positively regulates the pro-inflammatory function of macrophages

doi: 10.1007/s00018-025-05649-1

Figure Lengend Snippet: SDC3 deficient macrophages regulate EC migration and tube formation. ( a ) HUVEC migration over the indicated times in response to conditioned media (CM) from WT or SDC3 KO THP-1 macrophages stimulated with 100 ng/ml of IFNγ. Serum-free media (SFM) was used as negative control ( n = 4). ( b ) Quantification of HUVEC migration (Cell Index) shown in (a) at 48 h ( n = 4). ( c ) Representative images of Matrigel-grown HUVECs treated with supernatants from WT or SDC3 KO THP-1 macrophages stimulated with 100 ng/ml of IFNγ. Tube formation was imaged using an Olympus IX-83 inverted microscope (scale bar 1 mm, n = 4). ( d ) Quantification of total tube number shown in (c) at 24 h, relative to WT untreated control ( n = 4). ( e ) VEGFA quantification by ELISA in the supernatants of WT or SDC3 KO THP-1 macrophages stimulated with 100 ng/ml of IFNγ ( n = 4). ( f ) Angiogenic factor quantification in cell supernatants from WT or SDC3 KO THP-1 macrophages stimulated with 100 ng/ml of IFNγ, using the LEGENDplex™ HU Angiogenesis Panel 1 ( n = 4). Means ± SEM. Statistically significant difference from controls or between indicated groups is shown by ns: not significant, * p < 0.05 and *** p < 0.001

Article Snippet: SDC3 primary antibody (R&D Systems Cat. #: AF3539), was incubated overnight at 4 °C in blocking buffer.

Techniques: Migration, Negative Control, Inverted Microscopy, Control, Enzyme-linked Immunosorbent Assay

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Syndecan-3 positively regulates the pro-inflammatory function of macrophages

doi: 10.1007/s00018-025-05649-1

Figure Lengend Snippet: Macrophage-derived SDC3 plays a role in the regulation of the TME. The plethora of cells present in the TME interact to promote or inhibit tumour progression and growth. SDC3 is expressed by macrophages in response to the pro-inflammatory cytokine IFNγ and regulates many aspects of the TME. Firstly, SDC3 in macrophages reduces tumour proliferation and is necessary for macrophage-mediated tumour-cell phagocytosis. The expression of SDC3 promotes a pro-inflammatory phenotype in macrophages that results in the secretion of cytokines such as MCP-1 and IL-10, while reducing the release of IL-8. Additionally, SDC3 is necessary for the acquisition of an effector phenotype by T cells, shown by an increased expression of PD-1 and CD95 on T cells. Finally, macrophage-derived SDC3 plays a role in the inhibition of endothelial cell migration and proliferation through a reduction in the secretion of angiogenic factors such as VEGFA, PECAM-1 and IL-8

Article Snippet: SDC3 primary antibody (R&D Systems Cat. #: AF3539), was incubated overnight at 4 °C in blocking buffer.

Techniques: Derivative Assay, Expressing, Inhibition, Migration

Journal: Journal of Molecular Endocrinology

Article Title: Syndecans modulate ghrelin receptor signaling

doi: 10.1530/JME-24-0070

Figure Lengend Snippet: Syndecans (SDCs) impair β-arrestin2 recruitment at growth hormone secretagogue receptor (GHSR). (A) Effect of SDC1 on ghrelin-stimulated β-arrestin2–GHSR interaction in HEK293 cells. Values were corrected for Emax of the control (*** P < 0.0005 versus 1:0 ratio GHSR:SDC1; # P < 0.05, ## P < 0.005 versus 1:2 ratio GHSR:SDC1). (B) Effect of SDC1 on the kinetic profile of the β-arrestin2–GHSR interaction upon ghrelin stimulation (outcome of RM-ANOVA shown on graph: P T , effect of time; P Ratio , effect of GHSR:SDC ratio; P TxRatio , interaction. * P < 0.05, all other ratios versus 1:0 ratio GHSR:SDC1; # P < 0.05, 1:1 and 1:2 versus 1:0 ratio; $ P < 0.05, 1:2 versus 1:0 ratio). Data derived from curve fitting of the kinetic profiles for β-arrestin2 recruitment by GHSR in cells transfected with control or increasing amounts of SDC1 expression plasmid. (C) Initial rate of recruitment. (D) Magnitude of peak response (letters a and b represent groups that are significantly different from each other ( P < 0.05; one-way ANOVA). Dose–response curve (E) (*** P < 0.0005 versus control; # P < 0.05 versus 1:2 ratio GHSR:SDC1) and kinetic curve (F) of the effect of all SDCs (GHSR:SDC of 1:2) on β-arrestin2 recruitment at GHSR (outcome of RM-ANOVA shown on graph. * P < 0.05, *** P < 0.001, all SDCs versus control; # P < 0.05, SDC1, SDC2 and SDC4 versus control; $ P < 0.05, SDC2 and SDC4 versus control. P T , effect of time; P Ratio , effect of GHSR:SDC ratio; P TxRatio , interaction). Data derived from curve fitting of the kinetic profiles for β-arrestin2 recruitment by GHSR in cells transfected with control or SDC1, SDC2, SDC3 or SDC4 expression plasmids. (G) Initial rate of recruitment. (H) Magnitude of peak response (letters a and b represent groups that are significantly different from each other ( P < 0.05; one-way ANOVA). Data are presented as mean ± SEM of three independent experiments. Emax, maximum response.

Article Snippet: Because the SinoBiological SDC3 expression plasmid contained a 5′ truncated cDNA variant (SinoBiological, Germany; HG12158-UT; NCBI, BC013974), we generated a full-length pCMV3-SDC3 expression construct by inserting the correct 5′ region from a GenScript plasmid (GenScript, The Netherlands; Clone ID Ohu18173; NM_014654.4) utilizing Hind III to Kfl I restriction sites.

Techniques: Control, Derivative Assay, Transfection, Expressing, Plasmid Preparation