mouse anti chicken tnc  (Developmental Studies Hybridoma Bank)

Journal: eLife

Article Title: Postnatal mechanical loading drives adaptation of tissues primarily through modulation of the non-collagenous matrix

doi: 10.7554/eLife.58075

Figure Lengend Snippet: IFM and fascicle staining scores are shown for decorin (DCN), fibromodulin (FMOD), lubricin (PRG4), and tenascin-C (TNC) in the SDFT and CDET, alongside representative images of immunohistochemical staining in the postnatal SDFT. DCN and FMOD staining is found in both IFM (black triangle) and fascicle (white triangle). PRG4 staining in mainly located in the IFM (black triangle) and less staining can be found in the fascicle (white triangle). TNC staining is restricted to the IFM (black triangle) and absent from the fascicle (white triangle). A quantitative measure of elastin (ELN) is provided as percentage of wet weight, alongside a representative image of immunohistochemical staining in the postnatal SDFT. ELN staining is mainly located in the IFM (black triangle) and faint staining can be found in the fascicle (white triangle). Staining scores for elastin are provided in . ‡ significant change in tendon phase with development, ‡‡ significant interaction between tendon phase and development, * significant difference between tendons, a-d significant difference between age groups. Scale bar 100 µm. Error bars depict standard deviation. . Scoring of ELN staining for the IFM and fascicle in the SDFT and CDET through postnatal development.

Article Snippet: Primary antibodies were used at a concentration of 1:1500 for DCN (mouse IgG), 1:400 for FMOD (rabbit IgG), 1:200 for PRG4 (mouse IgG), 1:250 for TNC (mouse IgG, RRID: AB_785991 , Santa Cruz Biotechnology, Dallas, Texas), and 1:100 for ELN (mouse IgG, RRID: AB_2099589 , Abcam, Cambridge, UK).

Techniques: Staining, Immunohistochemical staining, Standard Deviation

Journal: BMC Cancer

Article Title: Neuropilin-1 promotes the oncogenic Tenascin-C/ integrin β3 pathway and modulates chemoresistance in breast cancer cells

doi: 10.1186/s12885-018-4446-y

Figure Lengend Snippet: Confirmation of DEGs from transcriptome analysis. a. KEGG pathway analysis of DEGs between BT-474 and BT-474 NRP-1 cells identified from RNA sequencing. Real-time qPCR was implemented on shortlisted DEGs obtained from RNA sequencing to confirm their differential expression. BT-474 NRP-1 cells significantly upregulated b. TNC expression and downregulated c, ACE, d, APOD , e, ATF3 , f, DDIT3 and g, P2RX6 expression. Cells transfected with the empty plasmid indicated similar expression to the BT-474 cells. Graphs represent the mean ± SEM of 3 independent experiments. Statistical analysis using independent samples t-test, p value < 0.05 considered as statistically significant ** p < 0.01, *** p < 0.001

Article Snippet: Here, 10 6 cells were seeded on a sterile positively charged slide, fixed with 4% Paraformaldehyde, permeabilized using 0.05% Triton X-100, followed by blocking with 5% goat serum and overnight incubation with a mixture of primary anti-rabbit NRP-1 antibody (1:100) (Abcam, UK) and anti-mouse TNC antibody (1:200) (Santa Cruz, USA) diluted in PBS.

Techniques: RNA Sequencing, Quantitative Proteomics, Expressing, Transfection, Plasmid Preparation

Journal: BMC Cancer

Article Title: Neuropilin-1 promotes the oncogenic Tenascin-C/ integrin β3 pathway and modulates chemoresistance in breast cancer cells

doi: 10.1186/s12885-018-4446-y

Figure Lengend Snippet: Tenascin C contributes to NRP-1 associated migration. a. Dual immunofluorescence staining (40× magnification scale bar 10 μm) of NRP-1 and TNC on BT-474 and BT-474 NRP-1 cells indicates their colocalization in the cytoplasm. Treatment of BT-474 NRP-1 cells with TNC targeted siRNA molecules, b , reduced TNC gene expression, c , reduced migratory capacity and d , downregulated NRP-1 and vimentin expression. (TNC protein was not detected on western blot due to the lack of specific antibody for this application.) The gene expression fold change was measured by comparing the basal levels detected in the empty plasmid transfected BT-474 or in the case of the siRNA experiment, to the control siRNA treated BT-474 and normalized to β-Actin and GUSB reference gene expression. Wound healing assay images (panel d, 5× magnification, scale bar 500 μm) taken on day 0 and day 2 after siRNA transfection. Graphs represent the mean ± SEM of three independent experiments. Statistical analysis using independent samples t-test, p-value < 0.05 considered as statistically significant. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Here, 10 6 cells were seeded on a sterile positively charged slide, fixed with 4% Paraformaldehyde, permeabilized using 0.05% Triton X-100, followed by blocking with 5% goat serum and overnight incubation with a mixture of primary anti-rabbit NRP-1 antibody (1:100) (Abcam, UK) and anti-mouse TNC antibody (1:200) (Santa Cruz, USA) diluted in PBS.

Techniques: Migration, Immunofluorescence, Staining, Gene Expression, Expressing, Western Blot, Plasmid Preparation, Transfection, Control, Wound Healing Assay

Journal: BMC Cancer

Article Title: Neuropilin-1 promotes the oncogenic Tenascin-C/ integrin β3 pathway and modulates chemoresistance in breast cancer cells

doi: 10.1186/s12885-018-4446-y

Figure Lengend Snippet: NRP-1 overexpression activates integrin β3 and TNFR2 pathways. Representative western blot images of protein lysates from untransfected BT-474, BT-474 NRP-1 and empty vector control cells blotted with indicated antibodies involved in a. Integrin signaling, and downstream signaling targets FAK, Akt, GSK3-β and NF-kB b. siRNA-mediated TNC downregulation decreased phosphorylation of FAK and Akt-473. c. Blots show levels of tumor necrosis factor receptors (TNFRs). GAPDH protein expression is indicated as a loading control. (The prefix P beside the antibody names indicates the phosphorylated form)

Article Snippet: Here, 10 6 cells were seeded on a sterile positively charged slide, fixed with 4% Paraformaldehyde, permeabilized using 0.05% Triton X-100, followed by blocking with 5% goat serum and overnight incubation with a mixture of primary anti-rabbit NRP-1 antibody (1:100) (Abcam, UK) and anti-mouse TNC antibody (1:200) (Santa Cruz, USA) diluted in PBS.

Techniques: Over Expression, Western Blot, Plasmid Preparation, Control, Phospho-proteomics, Expressing

Journal: BMC Cancer

Article Title: Neuropilin-1 promotes the oncogenic Tenascin-C/ integrin β3 pathway and modulates chemoresistance in breast cancer cells

doi: 10.1186/s12885-018-4446-y

Figure Lengend Snippet: NRP-1/TNC/Integrin β3/FAK/Akt/NF-kB schematic signaling pathway. Schema to illustrate the identified mechanism by which NRP-1 activates TNC-dependent integrin β3 signaling via phosphorylation of FAK/Akt-473/NF-κB p65. In addition, NRP-1 upregulates TNFR2 expression and downregulates BCRP/ABCG2, TNFR1, and PI3K/Akt-308. Solid black lines indicate identified direct interactions in the pathway whereas dashed lines indicate indirect effects of NRP-1 overexpression via unidentified mechanisms. The key denotes the arrows and colors used in the illustration

Article Snippet: Here, 10 6 cells were seeded on a sterile positively charged slide, fixed with 4% Paraformaldehyde, permeabilized using 0.05% Triton X-100, followed by blocking with 5% goat serum and overnight incubation with a mixture of primary anti-rabbit NRP-1 antibody (1:100) (Abcam, UK) and anti-mouse TNC antibody (1:200) (Santa Cruz, USA) diluted in PBS.

Techniques: Phospho-proteomics, Expressing, Over Expression