Journal: Cells

Article Title: Allogeneic Adipose-Derived Mesenchymal Stem Cell Transplantation Alleviates Atherosclerotic Plaque by Inhibiting Ox-LDL Uptake, Inflammatory Reaction and Endothelial Damage in Rabbits

doi: 10.3390/cells12151936

Figure Lengend Snippet: Detection of inflammatory cells and cytokines in aorta. Staining and analysis of macrophage marker molecules CD68, iNOS and Arginase-1 ( A ), inflammatory factors TNF-α and IL-6, and anti-inflammatory factors IL-10 and TGF-β ( B ): the levels of CD68, iNOS, IL-6 and TNF-α were lower, and the levels of IL-10 and TGF-β were higher in the T group compared with those in the model group, respectively (the areas of atherosclerotic plaque were circled with dashed lines, and indicated by green arrows) (n = 5–6 in each group). ** p < 0.01, bar = 100 µm.

Article Snippet: The cells were fixed in a 4% paraformaldehyde (E672002, Sangon Biotech), washed with PBS, blocked with serum for 30 min, incubated overnight with anti-arginase 1 (bs-23837R, Bioss) and iNOS (bs-0162R, Bioss) antibodies at 4 °C.

Techniques: Staining, Marker

Journal: Cells

Article Title: Allogeneic Adipose-Derived Mesenchymal Stem Cell Transplantation Alleviates Atherosclerotic Plaque by Inhibiting Ox-LDL Uptake, Inflammatory Reaction and Endothelial Damage in Rabbits

doi: 10.3390/cells12151936

Figure Lengend Snippet: ADSC and macrophage polarization assay in vitro. ( A – C ) Immunofluorescence detection of M1, M2 macrophage marker proteins iNOS and arginase-1. Compared with the model group, iNOS level was lower, and arginase-1 expression was higher in ADSC treatment group. Original magnification is 200. ( D ) ELISA results suggested that level of cytokine TNF-α secreted by M1 macrophages decreased in ADSCs group. ( E ) Western blot detection showed the expression of STAT6 signal molecules increased significantly in ADSCs group. * p < 0.05, ** p < 0.01.

Article Snippet: The cells were fixed in a 4% paraformaldehyde (E672002, Sangon Biotech), washed with PBS, blocked with serum for 30 min, incubated overnight with anti-arginase 1 (bs-23837R, Bioss) and iNOS (bs-0162R, Bioss) antibodies at 4 °C.

Techniques: In Vitro, Immunofluorescence, Marker, Expressing, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Cells

Article Title: Allogeneic Adipose-Derived Mesenchymal Stem Cell Transplantation Alleviates Atherosclerotic Plaque by Inhibiting Ox-LDL Uptake, Inflammatory Reaction and Endothelial Damage in Rabbits

doi: 10.3390/cells12151936

Figure Lengend Snippet: ADSC-derived exosomes and macrophage polarization assay in vitro. ( A ) TEM analysis presented that ADSC-derived exosomes were round membrane-bound vesicles and the diameter is about 30 mm to 150 nm as indicated by the red arrow. Scale bar is 200 nm. ( B ) Exosomes from rabbit ADSCs were identified by markers CD9, HSP70, TSG101 and Calnexin by Western blot. ( C , F ) Immunofluorescence detection of M1, M2 macrophage marker proteins iNOS and arginase-1. Compared with the model group, iNOS level was lower, and arginase-1 expression was higher in ADSC exosome treatment group. Original magnification is 200. ( D , E ) Western blot detection showed the expression of STAT6 signal molecules increased significantly in ADSC-derived exosome group. ( G ) ELISA results suggested that concentration of cytokine TNF-α decreased and IL-10 increased in ADSC exosomes group. * p < 0.05.

Article Snippet: The cells were fixed in a 4% paraformaldehyde (E672002, Sangon Biotech), washed with PBS, blocked with serum for 30 min, incubated overnight with anti-arginase 1 (bs-23837R, Bioss) and iNOS (bs-0162R, Bioss) antibodies at 4 °C.

Techniques: Derivative Assay, In Vitro, Western Blot, Immunofluorescence, Marker, Expressing, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Arthritis Research & Therapy

Article Title: Adiponectin associates with markers of cartilage degradation in osteoarthritis and induces production of proinflammatory and catabolic factors through mitogen-activated protein kinase pathways

doi: 10.1186/ar3512

Figure Lengend Snippet: Mitogen-activated protein kinase pathways are involved in adiponectin induced NO, IL-6, MMP-1 and MMP-3 production in osteoarthritis cartilage . The effects of mitogen-activated protein kinase inhibitors on adiponectin-induced NO production (A) , inducible nitric oxide synthase (iNOS) expression (B) , IL-6 production (C) , matrix metalloproteinase 1 (MMP-1) production (D) and MMP-3 production (E) in human osteoarthritis cartilage. Cartilage explants were incubated for 42 hours with adiponectin (1 μg/ml) and the inhibitor indicated. Samples were collected from six patients in (A) and (B) and from five patients in (C) through (E) . Results are expressed as means ± SEM. # P < 0.1, * P < 0.05, ** P < 0.01 and *** P < 0.001 compared to explants treated with adiponectin alone. PD98059 = extracellular signal-regulated kinase 1/2 (Erk1/2) inhibitor; SB220025 = p38 inhibitor; SP600125 = c-Jun N-terminal kinase (JNK) inhibitor. The inhibitor concentrations used were based on previous studies in our laboratory [ - ].

Article Snippet: Western blot analysis was performed as previously described [ ] using the following antibodies: rabbit anti-human pAb inducible nitric oxide synthase (iNOS), actin and c-Jun N-terminal kinase (JNK) antibodies, and horseradish-conjugated goat anti-rabbit immunoglobulin G antibody from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and rabbit anti-human pAb p38, phospho-p38, phospho-JNK, extracellular signal-regulated kinase 1/2 (Erk1/2) and phospho-Erk1/2 antibodies from Cell Signaling Technology, Inc (Beverly, MA, USA).

Techniques: Expressing, Incubation

Journal: Infection and Immunity

Article Title: Phospholipase C epsilon-1 (PLCƐ1) mediates macrophage activation and protection against tuberculosis

doi: 10.1128/iai.00495-23

Figure Lengend Snippet: PLCε1-/- macrophages display reduced anti-mycobacterial activity. (A) BMDMs prepared from B6 and PLCε1-/- mice were either infected with Mtb (for 6 days) or with Mtb mCherry (for 24 hours) at an MOI of 1; (B) bacterial burden was determined by plating cell lysate on 7H11 agar plates; (C and D) FACS analysis was performed to determine mCherry signal per cell; and (E) nitrate formation was estimated from the supernatant collected 24 hours post-Mtb mCherry infection in vitro; (n = 4–5). (F–H) Formalin-fixed paraffin-embedded lung sections from Mtb-infected mice were obtained at 50 dpi and subjected to IF for iNOS (green), CD206 (red), and F4/80 (white). Data represent mean ± SD, with a two-tailed unpaired t-test used to compare groups.

Article Snippet: Formalin-fixed paraffin-embedded (FFPE) lung sections were stained for iNOS (Rabbit anti-mouse/human, Novus Biologicals, Cat#NB300-605SS), CD206 (Goat anti-mouse, R&D systems, Cat# AF2535, RRID:AB_2063012), and F4/80 (Rat anti-mouse, BioRad, Clone Cl:A3-1, Cat#MCA497R, RRID:AB_323279).

Techniques: Activity Assay, Infection, In Vitro, Formalin-fixed Paraffin-Embedded, Two Tailed Test