Journal: Bioengineering

Article Title: Strontium Ranelate Inhibits Osteoclastogenesis through NF-κB-Pathway-Dependent Autophagy

doi: 10.3390/bioengineering10030365

Figure Lengend Snippet: Strontium ranelate suppressed osteoclast differentiation through autophagy in vitro. ( a ) TRAP staining images after pre-osteoclasts received different treatments for 5 days. ( b ) Monodansylcadaverine staining images after pre-osteoclasts received different treatments for 5 days. ( c ) Representative images of autophagosomes captured with a transmission electron microscope after pre-osteoclasts received different treatments for 5 days. Western blot assay and relative protein expression of the osteoclast markers TRAF6, c-Fos, MMP-9, MMP-14, and CTSK ( d ) and the autophagic proteins Beclin1, ATG5, LAMP2, LC3-I, LC3-II, and p62 ( e ) after pre-osteoclasts received different treatments for 5 days. (n = 3, mean ± SD, ** p < 0.01, *** p < 0.001).

Article Snippet: After that, the slices were incubated with a primary antibody at 4 °C overnight, including RANK (1:100; Zen), osteoprotegerin (OPG; 1:200; Servicebio), TRAF6 (1:100; Servicebio), nuclear factor of activated T cells 2 (NFATc2; 1:500; Servicebio), c-Fos (1:400; Servicebio), MMP-14 (1:100; Zen), CTSK (1:1000; Servicebio), p-IKK α/β (1:100; Affinity), IκBα (1:100; Zen), p65 (1:200; Zen), Beclin1 (1:200; Boster), LC3 (1:500; CST), LAMP2 (1:100; Zen), ATG5 (1:50; Zen), and p62 (1:200; Boster).

Techniques: In Vitro, Staining, Transmission Assay, Microscopy, Western Blot, Expressing

Journal: Bioengineering

Article Title: Strontium Ranelate Inhibits Osteoclastogenesis through NF-κB-Pathway-Dependent Autophagy

doi: 10.3390/bioengineering10030365

Figure Lengend Snippet: Strontium ranelate inhibited osteoclast differentiation through NF-κB-pathway-dependent autophagy in vitro. ( a ) TRAP staining images after pre-osteoclasts received different treatments for 5 days. ( b ) Monodansylcadaverine staining images after pre-osteoclasts received different treatments for 5 days. ( c ) Representative images of autophagosomes captured with a transmission electron microscope after pre-osteoclasts received different treatments for 5 days. Western blot assay and relative protein expression of the osteoclast marker CTSK, the proteins central to the NF-κB pathway (p-IKKα/β, IκBα, and p65) ( d ), and the autophagic proteins Beclin-1, ATG5, LAMP2, LC3-I, LC3-II, and p62 ( e ) after pre-osteoclasts were treated with Bay 11-7082 for 5 days. ( f ) Western blot assay and relative protein expression of the proteins central to the NF-κB pathway after pre-osteoclasts were treated with SR for 5 days. (n = 3, mean ± SD, *** p < 0.001).

Article Snippet: After that, the slices were incubated with a primary antibody at 4 °C overnight, including RANK (1:100; Zen), osteoprotegerin (OPG; 1:200; Servicebio), TRAF6 (1:100; Servicebio), nuclear factor of activated T cells 2 (NFATc2; 1:500; Servicebio), c-Fos (1:400; Servicebio), MMP-14 (1:100; Zen), CTSK (1:1000; Servicebio), p-IKK α/β (1:100; Affinity), IκBα (1:100; Zen), p65 (1:200; Zen), Beclin1 (1:200; Boster), LC3 (1:500; CST), LAMP2 (1:100; Zen), ATG5 (1:50; Zen), and p62 (1:200; Boster).

Techniques: In Vitro, Staining, Transmission Assay, Microscopy, Western Blot, Expressing, Marker

Journal: Bioengineering

Article Title: Strontium Ranelate Inhibits Osteoclastogenesis through NF-κB-Pathway-Dependent Autophagy

doi: 10.3390/bioengineering10030365

Figure Lengend Snippet: Strontium ranelate regulated the expression of autophagic proteins in rats. ( a ) Immunohistochemical staining images of ATG5, Beclin1, LAMP2, LC3, and p62 at the pressure side of the first molars on day 7 in different groups. ( b ) The mean optical density (MOD) of ATG5, Beclin1, LAMP2, LC3, and p62 at the pressure side of the first molars on days 3, 7, and 14 in different groups. (n = 5, mean ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001). ( c ) Immunofluorescence staining images of ATG5, Beclin1, LAMP2, LC3, and p62 at the pressure side of the first molars on day 7 in different groups. The white dashed lines indicate the margin of the tooth root.

Article Snippet: After that, the slices were incubated with a primary antibody at 4 °C overnight, including RANK (1:100; Zen), osteoprotegerin (OPG; 1:200; Servicebio), TRAF6 (1:100; Servicebio), nuclear factor of activated T cells 2 (NFATc2; 1:500; Servicebio), c-Fos (1:400; Servicebio), MMP-14 (1:100; Zen), CTSK (1:1000; Servicebio), p-IKK α/β (1:100; Affinity), IκBα (1:100; Zen), p65 (1:200; Zen), Beclin1 (1:200; Boster), LC3 (1:500; CST), LAMP2 (1:100; Zen), ATG5 (1:50; Zen), and p62 (1:200; Boster).

Techniques: Expressing, Immunohistochemical staining, Staining, Immunofluorescence

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Genome-wide differential genetic profiling characterizes colorectal cancers with genetic instability and specific routes to HLA class I loss and immune escape

doi: 10.1007/s00262-011-1147-7

Figure Lengend Snippet: List of inflammatory- and macrophage markers-related gene-specific probes differentially expressed ( P < 0.05) between MSI and MSS/HLA-positive (control) colorectal cancers

Article Snippet: The following mouse mAbs were used to stain tumour infiltrates: GRT2 (anti-CD45), produced in our lab [ 31 ]; OKT3 (anti-CD3 hybridoma, ATCC, Teddington, UK), OKT8 (anti-CD8 hybridoma, ATCC), anti-CD4 (Clone RPA-T4, Becton–Dickinson Biosciences (BDB), San Jose, CA), anti-perforin (Clone δG9, BD Biosciences), anti-CD64 (Clone 10.1, BD Biosciences), anti-CD206 (macrophage mannose receptor; Clone 19.2, BD Biosciences), anti-CD163 (macrophage scavenger receptor; Clone Ber-Mac3, MBL, Woburn, MA) and anti-CD56 (Clone 123C3, Dako, Barcelona, Spain.

Techniques: Significance Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Genome-wide differential genetic profiling characterizes colorectal cancers with genetic instability and specific routes to HLA class I loss and immune escape

doi: 10.1007/s00262-011-1147-7

Figure Lengend Snippet: Immunohistological results of microarray data validation in CRC tissue samples. a MSI tumour (CRC-6), b MSS/HLA-positive (control) tumour (CRC-19) and c MSS/HLA-negative tumour (CRC-14). A higher infiltration by CD8+ cells was observed in MSI tumour versus control CRC, whereas no CD8+ cells were observed in MSS/HLA-negative CRC. d Elevated expression of CD3 complex ζ-chain in MSI tumour (CRC-1) compared with e Control tumour (CRC-19) and f MSS/HLA-negative sample (CRC-14). Perforin staining in paraffin-embedded samples was higher in g MSI tumours (CRC-6) than in h Control tumours (CRC-16) and i MSS/HLA-negative CRCs (CRC-11). No differences were observed in infiltration by M1 (CD64) and M2 (CD163) macrophages in CRC samples. j, m MSI tumour (CRC-1); k, n MSS/HLA-positive sample (CRC-19); l MSS/HLA-negative CRCs (CRC-11) and o (CRC-12)

Article Snippet: The following mouse mAbs were used to stain tumour infiltrates: GRT2 (anti-CD45), produced in our lab [ 31 ]; OKT3 (anti-CD3 hybridoma, ATCC, Teddington, UK), OKT8 (anti-CD8 hybridoma, ATCC), anti-CD4 (Clone RPA-T4, Becton–Dickinson Biosciences (BDB), San Jose, CA), anti-perforin (Clone δG9, BD Biosciences), anti-CD64 (Clone 10.1, BD Biosciences), anti-CD206 (macrophage mannose receptor; Clone 19.2, BD Biosciences), anti-CD163 (macrophage scavenger receptor; Clone Ber-Mac3, MBL, Woburn, MA) and anti-CD56 (Clone 123C3, Dako, Barcelona, Spain.

Techniques: Microarray, Positive Control, Expressing, Staining

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Genome-wide differential genetic profiling characterizes colorectal cancers with genetic instability and specific routes to HLA class I loss and immune escape

doi: 10.1007/s00262-011-1147-7

Figure Lengend Snippet: Summarized results of immunohistochemical study of leucocyte infiltration in CRC samples

Article Snippet: The following mouse mAbs were used to stain tumour infiltrates: GRT2 (anti-CD45), produced in our lab [ 31 ]; OKT3 (anti-CD3 hybridoma, ATCC, Teddington, UK), OKT8 (anti-CD8 hybridoma, ATCC), anti-CD4 (Clone RPA-T4, Becton–Dickinson Biosciences (BDB), San Jose, CA), anti-perforin (Clone δG9, BD Biosciences), anti-CD64 (Clone 10.1, BD Biosciences), anti-CD206 (macrophage mannose receptor; Clone 19.2, BD Biosciences), anti-CD163 (macrophage scavenger receptor; Clone Ber-Mac3, MBL, Woburn, MA) and anti-CD56 (Clone 123C3, Dako, Barcelona, Spain.

Techniques: Immunohistochemical staining

Journal: Journal of Thrombosis and Haemostasis

Article Title: Off‐target effects of oral anticoagulants – vascular effects of vitamin K antagonist and non‐vitamin K antagonist oral anticoagulant dabigatran etexilate

doi: 10.1111/jth.15289

Figure Lengend Snippet: Short‐term oral anticoagulant effects on atherosclerotic lesions. (A) Schematic overview of experimental setup of the animal experiment. (B) Apoe −/− mice were treated for 6 weeks with WTD without or with warfarin or dabigatran, after which animals were sacrificed. Aortic arches of Apoe −/− mice on WTD without or with warfarin or dabigatran showed no significant difference on plaque size at 6 weeks of treatment. (C) MAC3+ cells were significantly increased in lesions of warfarin‐treated mice compared with control ( p < .05) and dabigatran ( p < .01). (D) Six weeks of warfarin treatment significantly increased ucMGP in atherosclerosis lesions compared to control and dabigatran. a, adventitia; d, aorta descendens; h, heart; i, intima; l, lumen; m, media; ucMGP, uncarboxylated matrix Gla protein; WTD, Western type diet

Article Snippet: The following antibodies were used: anti‐uncarboxylated matrix Gla‐protein (1:400; IDS, Boldon, UK), anti‐bone morphogenetic protein 4 (BMP4; 1:50; Santa Cruz Biotechnology, Dallas, TX), anti‐8‐hydroxyguanosine (8OHdG; 1:300; Meridian Life Science, TN), anti‐MAC3 (1:50, Bioscience, Merck KGaA), and anti‐integrin ß 3 (ITGB3 1:100, Sigma‐Aldrich).

Techniques: Western Blot