Journal: Scientific Reports

Article Title: Inhibition of NUCKS Facilitates Corneal Recovery Following Alkali Burn

doi: 10.1038/srep41224

Figure Lengend Snippet: ( A ) (i) Corneal epithelial cells isolated from NWT and NKO mice were treated with LPS in vitro ; expression of PIκB was measured (ii) Graphic representation for PIκB data in (i) is shown. ( B ) (i) Corneal epithelial cells were treated with LPS or LPS + NFκB inhibitor, SC 154. Expression of PIκB and TNFα was tested. (ii and iii) Graphic representation of the relative expression of (ii) PIκB and (iii) TNFα, respectively in (i) is shown. ( C ) On treatment with LPS, expression of various inflammatory cytokines (IL6, IL10, INFγ, IP10, TNFα and IL12) was compared and analyzed by flow cytometry analysis. Results for (i) NWT group and (ii) NKO group are shown.

Article Snippet: Specific antibodies for PIκb and TNFα (1:1000 abcam) were used.

Techniques: Isolation, In Vitro, Expressing, Flow Cytometry

Journal: Best practice & research. Clinical gastroenterology

Article Title: Therapeutic approaches for celiac disease

doi: 10.1016/j.bpg.2015.04.005

Figure Lengend Snippet: Potential therapies for celiac disease.

Article Snippet: Anti-TNF-α , Adalimumab , Monoclonal antibody , AbbVie, USA , Approved.

Techniques: Genetically Modified

Journal: Nanomaterials

Article Title: Few Layer Graphene Does Not Affect Cellular Homeostasis of Mouse Macrophages

doi: 10.3390/nano10020228

Figure Lengend Snippet: Cytokine secretion by BMDMs. ( A ) IL-6, ( B ) TNF-α, and ( C ) IL-1β levels in cell supernatant after treatment with or without FLG (3, 10, 30, 100 µg/mL), and LPS (1 µg/mL) for 24 h. A One-way ANOVA followed by Bonferroni’s post-test was performed to determine the statistical differences for control untreated cells versus FLG-treated samples and LPS-treated cells (* p < 0.05).

Article Snippet: In short, polyvinyl microtiter 96-well plates (Falcon) were coated overnight at 4 °C with 50 µL/well of purified rat anti-mouse IL-6 (0.5 mg/mL, BD Pharmingen #554400), rabbit anti-mouse TNF-α (0.5 mg/mL, BD Pharmingen #557516) and IL-1β (1 mg/mL, BD Biosciences, Le Pont de Claix, France), #51_26661E), diluted in coating buffer (Carbonate/bicarbonate buffer 0.05 M, pH 9.6).

Techniques:

Journal: bioRxiv

Article Title: CD95L concatemers highlight difference in the manner CD95-mediated apoptotic and non-apoptotic pathways are triggered

doi: 10.1101/2023.06.22.546070

Figure Lengend Snippet: A . The amino acid residues 103 to 143 were missing in the crystal structure. Their structure was built through extensive molecular dynamic simulations. The final state of the reconstituted CD95L trimeric model with Nt region in yellow is depicted. B . Addition of the amino acid residues 103 to 143 to the crystalized CD95L did not affect the experimental 3D structure of the protomer. C . Based on the reconstituted CD95L trimeric model showed in Figure A, a molecular modeling approach was applied to predict the minimal repetition of the flexible linker GGGGS (in blue) to connect covalently three CD95L protomers without affecting the global structure of the crystalized homo-trimeric CD95L. D . Schematic representation of the indicated concatemers. E . The CD95L concatemer-encoding pcDNA3 vectors and empty or human full length CD95L-encoding pLVX-IRES-tdTomato vectors were transfected in HEK/293T cells and supernatants were harvested after 7 days. After ultracentrifugation to eliminate exosomes, supernatants were resolved in reducing and denaturing conditions (SDS-PAGE). Indicated immunoblotting was performed. F . The molecular weights of the different CD95L were assessed in native condition using BN-PAGE method.

Article Snippet: Anti-CD95L antibody (G247-4, #556387) was acquired from BD Bioscience.

Techniques: Transfection, SDS Page, Western Blot

Journal: bioRxiv

Article Title: CD95L concatemers highlight difference in the manner CD95-mediated apoptotic and non-apoptotic pathways are triggered

doi: 10.1101/2023.06.22.546070

Figure Lengend Snippet: A . CD95L constructs secreted by HEK/293T cells were dosed by ELISA and their cytotoxic activity was evaluated by MTS assay using the CD95 sensitive cell line Jurkat. EC 50 was assessed for the monomeric CD95L and the indicated concatemers. Data represent means and SD of three independently performed experiments. B . The cytotoxic activity of CD95L constructs was evaluated by MTS assay using the CD95 sensitive cell line Jurkat and its FADD-deficient counterpart, namely I2.1 cell line. Data represent means and SD of three independently performed experiments.

Article Snippet: Anti-CD95L antibody (G247-4, #556387) was acquired from BD Bioscience.

Techniques: Construct, Enzyme-linked Immunosorbent Assay, Activity Assay, MTS Assay

Journal: bioRxiv

Article Title: CD95L concatemers highlight difference in the manner CD95-mediated apoptotic and non-apoptotic pathways are triggered

doi: 10.1101/2023.06.22.546070

Figure Lengend Snippet: A . The cytosolic calcium concentration ([Ca 2+ ] cyt ) in Jurkat cells exposed to the indicated concentration of CD95L was assessed using fura PE3-AM. Ratio values (R) were normalized to pre-stimulated values (R0) to yield R/R0 values (relative [Ca2+]). Data represent the mean □ ± □ SD. B . Same experiment as A with the metalloprotease-cleaved CD95L. C . Left panel: area under the curves (AUC) of the Ca 2+ responses obtained in A and B are represented. Right panel: AUC of the Ca 2+ response obtained in Jurkat cells exposed to the higher concentration of ligands (100 ng/mL).

Article Snippet: Anti-CD95L antibody (G247-4, #556387) was acquired from BD Bioscience.

Techniques: Concentration Assay

Journal: BioMed Research International

Article Title: Imbalance of the Nerve Growth Factor and Its Precursor as a Potential Biomarker for Diabetic Retinopathy

doi: 10.1155/2015/571456

Figure Lengend Snippet: Shedding of p75 NTR is consistent in vitreous and serum. (a) Representative bands show p75 NTR expression in vitreous and serum for diabetic compared to nondiabetic control groups. The full length p75 NTR (75 kD) and receptor ectodomain (50 kD) had similar levels of expression in control and diabetic (DB) groups of both vitreous and serum. The possible proteolytic C terminal fragment (CTF) and intracellular domain (ICD) appeared at 27 kD and 22 kD. Differences in expression patterns between vitreous and serum as well as between diabetic and control groups were evident for both CTF and ICD. (b) In vitreous, 27 kD p75 NTR receptor fragment was significantly increased in diabetic (1.65-fold ± 0.23) compared to nondiabetic control group ( N = 4–11, * P < 0.05). (c) In serum, a significant increase in 22 kD p75 NTR occurred in diabetic samples (1.85-fold ± 0.30) compared to nondiabetic controls ( N = 6–10, * P < 0.05).

Article Snippet: The following antibodies were used for immunoblotting: rabbit polyclonal anti-NGF and anti-proNGF (Alomone Labs, Israel) and rabbit polyclonal p75 NTR (1 : 5000), a kind gift from Dr. Bruce Carter, Vanderbilt University, Nashville, TN.

Techniques: Expressing

Journal: BioMed Research International

Article Title: Imbalance of the Nerve Growth Factor and Its Precursor as a Potential Biomarker for Diabetic Retinopathy

doi: 10.1155/2015/571456

Figure Lengend Snippet: Expression of p75 NTR receptor is consistent in vitreous and serum. Results are shown for p75 NTR expression in vitreous and serum of diabetic (DB) and control participants normalized to Ponceau S and respective controls. Full length p75 NTR receptor (75 kD) was not significantly different in diabetic sample compared to control groups in either (a) vitreous ( N = 4–11) or (b) serum ( N = 6–9). The p75 NTR ectodomain (50 kD) was also not significantly different in diabetic compared to control groups in (c) vitreous ( N = 4–11) or (d) serum ( N = 6–9).

Article Snippet: The following antibodies were used for immunoblotting: rabbit polyclonal anti-NGF and anti-proNGF (Alomone Labs, Israel) and rabbit polyclonal p75 NTR (1 : 5000), a kind gift from Dr. Bruce Carter, Vanderbilt University, Nashville, TN.

Techniques: Expressing