Journal: PLoS ONE

Article Title: A Ribosomal S-6 Kinase–Mediated Signal to C/EBP-β Is Critical for the Development of Liver Fibrosis

doi: 10.1371/journal.pone.0001372

Figure Lengend Snippet: A. TRADD, C/EBPβ, RIP and caspase 8 immunoblots were performed on C/EBPβ immunoprecipitates from primary human HSC treated with an ERK1/2 inhibitor (10 µM) or the cell permeant Ac-KA217VD-CHO peptide (200 µM). Blocking the phosphorylation of C/EBPβ by RSK with the ERK1/2 inhibitor or the cell permeant Ac-KA217VD-CHO (KAVD) peptide, increased the association between C/EBPβ, active caspase 8, TRADD and RIP. β-Actin was used as an internal control for the immunoprecipitations. B. TNFR1 and C/EBPβ immunoblots were performed on C/EBPβ immunoprecipitates from HSC isolated from mice treated with CCl 4 for 12 or 16 weeks as described in . Blocking the phosphorylation of C/EBPβ by RSK with C/EBPβ-Ala217 transgene or the cell permeant Ac-KA217VD-CHO peptide increased the association between C/EBPβ and TNFR1. β-Actin was used as an internal control for the immunoprecipitations. C. TRAF2 and C/EBPβ immunoblots were performed on C/EBPβ immunoprecipitates from HSC isolated from mice treated with CCl 4 for 12 or 16 weeks as described in . Blocking the phosphorylation of C/EBPβ by RSK with C/EBPβ-Ala217 transgene or the cell permeant Ac-KA217VD-CHO peptide increased the association between C/EBPβ and TRAF2. β-Actin was used as an internal control for the immunoprecipitations. D. Cytochrome C and Apaf1 immunoblots were performed on cytochrome C immunoprecipitates in livers from mice treated with CCl 4 for 12 or 16 weeks as described in . Blocking the phosphorylation of C/EBPβ by RSK with C/EBPβ-Ala217 transgene or the cell permeant Ac-KA217VD-CHO peptide increased the association between cytochrome C and Apaf1. β-Actin was used as an internal control for the immunoprecipitations.

Article Snippet: Pre-cleared stellate cell lysates were incubated for 2 h with purified C/EBPβ, RSK, TRADD or caspase 8 antibodies followed by the addition of A/G+ agarose (Santa Cruz Biotechnology) for 12 h. The immunoprecipitation reactions each contained 500 μg of total protein and 2 μg antibody (or purified IgG pre-immune serum as negative control).

Techniques: Western Blot, Blocking Assay, Phospho-proteomics, Control, Isolation

Journal: Antioxidants

Article Title: Anti-Amnesic Effect of Walnut via the Regulation of BBB Function and Neuro-Inflammation in Aβ 1-42 -Induced Mice

doi: 10.3390/antiox9100976

Figure Lengend Snippet: List of antibodies and their information used in this study.

Article Snippet: TNFR1 , CSB-PA621879EA01HU , 1:1000 , Cusabio (Hubei, China).

Techniques:

Journal: Antioxidants

Article Title: Anti-Amnesic Effect of Walnut via the Regulation of BBB Function and Neuro-Inflammation in Aβ 1-42 -Induced Mice

doi: 10.3390/antiox9100976

Figure Lengend Snippet: Protective effect of walnut ( Juglans regia L.) extract on Aβ-induced neuro-inflammation: ( A ) protein expression levels; ( B ) representative Western blots for total protein and expression of tumor necrosis factor-alpha (TNF-α) ( B ), tumor necrosis factor receptor 1 (TNFR1) ( C ), phosphorylated c-Jun N-terminal kinase (p-JNK) ( D ), phosphorylated nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor (p-IκB) ( E ), cyclooxygenase-2 (COX-2) ( F ), and interleukin 1 beta (IL-1β) ( G ) in mice brain tissues. Results shown are means ± SD ( n = 3). Data are statistically represented at * = significantly different from the NC group; # = significantly different from Ab group, respectively; * and # p < 0.05.

Article Snippet: TNFR1 , CSB-PA621879EA01HU , 1:1000 , Cusabio (Hubei, China).

Techniques: Expressing, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: RIPK1 and TRADD Regulate TNF-Induced Signaling and Ripoptosome Formation

doi: 10.3390/ijms222212459

Figure Lengend Snippet: RIPK1 prevented TNF-dependent TRADD modification and degradation independent of cIAP and could be prevented by A20 over-expression. Control cells and RIPK1 KO clones were treated ( A ) with TNF for the indicated time points. ( B ) The cells from ( A ) were pretreated with BTZ and CHX for 5 h before stimulation with TNF for the indicated time points. ( C ) The cells from ( A ) were pretreated with IAP antagonist before stimulation with TNF for 2 h. ( D ) Control and RIPK1 KO cells were transduced with control or A20 containing LV and stimulated with TNF for the indicated time points. Protein expression was analyzed by WB. The WB shown are representative of at least two independent experiments.

Article Snippet: The following antibodies (Abs) were used for WB: RIPK1 (R41220), TRADD (T50320), and FADD (F36620) Abs were purchased from BD Transduction Laboratories, San Diego, California.

Techniques: Modification, Over Expression, Clone Assay, Transduction, Expressing

Journal: International Journal of Molecular Sciences

Article Title: RIPK1 and TRADD Regulate TNF-Induced Signaling and Ripoptosome Formation

doi: 10.3390/ijms222212459

Figure Lengend Snippet: Suggested model of TRADD and RIPK1 functions in TNF signaling and ripoptosome formation. Under normal conditions (yellow field), the signal initiated by TNF through TNF-R1 proceeds via the assembly of TNF complex I and initiates NF-κB and MAPK signaling. Assembly of complex I and stabilization of NIK are the requirements for ripoptosome formation, which can direct the cell to apoptosis or necroptosis. NIK stabilization can be blocked by cIAPs or TRADD. When RIPK1 is missing (green field) complex I is assembled but NF-κB and MAPK signaling are partially blocked. No ripoptosome is formed. The assembly of pseudocomplex is unable to direct the cell-to-cell death. Unknown E3 ubiquitin ligase can ubiquitinate TRADD and direct it to proteasomal degradation. When TRADD is missing (red field), complex I consists only of unmodified RIPK1 and both NF-κB and MAPK signaling are partially blocked. NIK stabilization is simplified and ripoptosome is assembled and can direct the cell to apoptosis or necroptosis.

Article Snippet: The following antibodies (Abs) were used for WB: RIPK1 (R41220), TRADD (T50320), and FADD (F36620) Abs were purchased from BD Transduction Laboratories, San Diego, California.

Techniques: