Journal: International Journal of Molecular Medicine

Article Title: Histone deacetylase-4 and histone deacetylase-8 regulate interleukin-1β-induced cartilage catabolic degradation through MAPK/JNK and ERK pathways

doi: 10.3892/ijmm.2018.3410

Figure Lengend Snippet: IL-1β promoted the expression of ADAMTS-4 and ADAMTS-5 through the activation of MAPK signaling. (A) Of the 3 MAPK isoforms, ERK1/2 and JNK were markedly phosphorylated following treatment with IL-1β. The gene expression of (B) ADAMTS-5 and (C) ADAMTS-4 was facilitated by C6 (ERK activator) or Ani (JNK activator). The IL-1β-induced gene expression of (B) ADAMTS-5 and (C) ADAMTS-4 was inhibited by FR (ERK inhibitor) or SP (JNK inhibitor). Data present the means ± standard deviation, n=3. * P<0.05. IL, interleukin; ADAMTS, a disintegrin and metalloproteinase with thrombospondin motifs; MAPK, mitogen-activated protein kinase; ERK, extracellular signal-regulated kinase; JNK, c-Jun N-terminal kinase; DMSO, dimethyl sulfoxide; C6, Ceramide C6; Ani, Anisomycin; FR, FR180204; SP, SP600125; p, phosphorylated; t, total.

Article Snippet: For the kinase assays, rACs were pretreated with specific inhibitors of ERK (FR180204 at 50 μ M) and JNK (SP600125 at 10 μ M; both from Selleck Chemicals, Houston, TX, USA) and exposed to IL-1β, or treated with the specific activators of ERK (ceramide C6 at 50 μ M; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) or JNK (anisomycin at 1 μ M; Selleck Chemicals) alone. rACs treated with DMSO were used as a vehicle control and with IL-1β were used as a positive control.

Techniques: Expressing, Activation Assay, Gene Expression, Standard Deviation

Journal: International Journal of Molecular Medicine

Article Title: Histone deacetylase-4 and histone deacetylase-8 regulate interleukin-1β-induced cartilage catabolic degradation through MAPK/JNK and ERK pathways

doi: 10.3892/ijmm.2018.3410

Figure Lengend Snippet: HDAC regulates the osteoarthritis-related genes and protein expression of rACs via ERK1/2 and JNK. (A) IL-1β-induced ERK1/2 phosphorylation was blocked by TSA, and not blocked by PCI; JNK phosphorylation was blocked by TSA and PCI. (B) HDAC inhibitors (TSA or PCI) suppressed HDAC expression at both mRNA and protein level, and prevented the upregulation of osteoarthritis-related genes (col X, COX2, ADAMTS-5 and ADAMTS-4) expression induced by IL-1β. Results are presented with respect to GAPDH expression. (C) siRNAs (targeting HDAC4 or HDAC8) suppressed HDAC expression at both mRNA and protein level, and prevented the upregulation of osteoarthritis-related gene expression induced by IL-1β. Results are expressed with respect to GAPDH expression. Data represent the means ± standard deviation, n=3. * P<0.05. HDAC, histone deacetylase; ERK, extracellular signal-regulated kinase; JNK, c-Jun N-terminal kinase; IL, interleukin; TSA, Trichostatin A; DMSO, dimethyl sulfoxide; PCI, PCI-34051; col X, type X collagen; COX2, cyclooxygenase; ADAMTS, a disintegrin and metalloproteinase with thrombospondin motifs; siRNA, small interfering RNA; p, phosphorylated; t, total. rACs, rat articular chondrocytes.

Article Snippet: For the kinase assays, rACs were pretreated with specific inhibitors of ERK (FR180204 at 50 μ M) and JNK (SP600125 at 10 μ M; both from Selleck Chemicals, Houston, TX, USA) and exposed to IL-1β, or treated with the specific activators of ERK (ceramide C6 at 50 μ M; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) or JNK (anisomycin at 1 μ M; Selleck Chemicals) alone. rACs treated with DMSO were used as a vehicle control and with IL-1β were used as a positive control.

Techniques: Expressing, Phospho-proteomics, Gene Expression, Standard Deviation, Histone Deacetylase Assay, Small Interfering RNA

Journal: International Journal of Molecular Medicine

Article Title: Histone deacetylase-4 and histone deacetylase-8 regulate interleukin-1β-induced cartilage catabolic degradation through MAPK/JNK and ERK pathways

doi: 10.3892/ijmm.2018.3410

Figure Lengend Snippet: Possible signaling pathways associated with ADAMTS, col X, and COX-2 regulation by IL-1β in chondrocytes. ADAMTS, a disintegrin and metalloproteinase with thrombospondin motifs; col X, type X collagen; COX2, cyclooxygenase; IL, interleukin; HDAC, histone deacetylase; ERK, extracellular signal-regulated kinase; JNK, c-Jun N-terminal kinase; IL, interleukin; TSA, Trichostatin A; PCI, PCI-34051.

Article Snippet: For the kinase assays, rACs were pretreated with specific inhibitors of ERK (FR180204 at 50 μ M) and JNK (SP600125 at 10 μ M; both from Selleck Chemicals, Houston, TX, USA) and exposed to IL-1β, or treated with the specific activators of ERK (ceramide C6 at 50 μ M; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) or JNK (anisomycin at 1 μ M; Selleck Chemicals) alone. rACs treated with DMSO were used as a vehicle control and with IL-1β were used as a positive control.

Techniques: Protein-Protein interactions, Histone Deacetylase Assay

Journal: Frontiers in Immunology

Article Title: Complement membrane attack complex is an immunometabolic regulator of NLRP3 activation and IL-18 secretion in human macrophages

doi: 10.3389/fimmu.2022.918551

Figure Lengend Snippet: Reagents List.

Article Snippet: Ruthenylated anti-C6 , (Quidel, ruthenylated in-house) , A219.

Techniques: Purification, Recombinant, In Vivo, Generated, Membrane, Enzyme-linked Immunosorbent Assay, Sequencing, Software

Journal: PLoS pathogens

Article Title: Exploitation of the complement system by oncogenic Kaposi's sarcoma-associated herpesvirus for cell survival and persistent infection.

doi: 10.1371/journal.ppat.1004412

Figure Lengend Snippet: Figure 7. Activation of complement pathway induces STAT3 phosphorylation to enhance cell survival of latently KSHV-infected endothelial cells. (A) The enhanced cell survival of KSHV-infected endothelial cells by complement was mediated by the STAT3 pathway. TIME- KSHV cells were cultured for 48 h in normal human serum in growth factor-depleted medium with and without JAK or STAT3 inhibitor, and the numbers of dead cells (left panel) and live cells (right panel) were determined. Cells cultured in heat-inactivated human serum were used as controls. Results are means 6 SD from three independent experiments with three repeats. * P,0.05, ** P,0.01 and *** P,0.001 by Student’s t-test. (B) Complement activation induced STAT3 tyrosine phosphorylation in TIME-KSHV cells. STAT3 tyrosine (Y705) and serine (S727) phosphorylation in TIME and TIME-KSHV cells switched from full endothelial cell medium with growth factors and 10% heat-inactivated human serum to endothelial cell medium depleted of growth factors with 10% heat-inactivated or normal human serum for the specified lengths of time. (C) Complement activation was required for the enhanced STAT3 tyrosine phosphorylation of TIME-KSHV cells. STAT3 tyrosine phosphorylation was examined in cells cultured for 24 h in 10% C3-depleted human serum or C3-depleted human serum reconstituted with purified C3 protein in endothelial cell medium depleted of growth factors. (D) Formation of C5b-9 complexes was required for the enhanced STAT3 tyrosine phosphorylation. STAT3 tyrosine phosphorylation was examined in cells cultured for 24 h in 10% C6-depleted human serum or C6-depleted human serum reconstituted with purified C6 protein in endothelial cell medium deprived of growth factors. (E) STAT3 tyrosine phosphorylation was unchanged without the continuous presence of normal human serum and growth factors. STAT3 tyrosine phosphorylation was examined in cells cultured in 10% heat-inactivated or normal human serum for 1 h, and then in endothelial cell medium without any serum and growth factors for additional 23 h. (F) JAK but not Src activation mediated STAT3 tyrosine phosphorylation. STAT3 tyrosine phosphorylations was examined in TIME-KSHV cells cultured in 10% normal human serum in endothelial cell medium depleted of growth factors for 24 h with or without the presence of STAT3, JAK or Src inhibitor. doi:10.1371/journal.ppat.1004412.g007

Article Snippet: C1q-depleted human serum, C3-depleted human serum, C6-depleted human serum, factor B-depleted human serum, purified C3 protein, and purified C6 protein were purchased from Quidel Corporation (San Diego, CA).

Techniques: Activation Assay, Phospho-proteomics, Infection, Cell Culture, Purification