Journal: Oncology Letters

Article Title: Significance of interstitial tumor-associated macrophages in the progression of lung adenocarcinoma

doi: 10.3892/ol.2016.5270

Figure Lengend Snippet: Typical expression images of immunochemistry staining in tissue microarrays. (A and B) Low expression of CD68 in MIA tissues. Original magnification, (A) ×100 and (B) ×200. (C and D) High expression of CD68 in LPA tissues. Original magnification, (C) ×100 and (D) ×200. (E) Negative expression of IL-6 in MIA tissues. Original magnification, ×200. (F) Positive expression of IL-6 in LPA tissues. Original magnification, ×200. (G) Negative expression of CSF-1 in MIA tissues. Original magnification, ×200. (H) Positive expression of CSF-1 in LPA tissues. Original magnification, ×200. (I) High E-cadherin expression in MIA tissues. Original magnification, ×100. (J) Low E-cadherin expression in LPA tissues. Original magnification, ×100. (K) High Snail expression in LPA tissues. Original magnification, ×100. (L) High MMP-2 expression in LPA tissues. Original magnification, ×100. CD, cluster of differentiation; MIA, minimally invasive adenocarcinoma; LPA, lepidic predominant adenocarcinoma; CSF, colony-stimulating factor; IL, interleukin; MMP, metalloproteinase.

Article Snippet: Immunohistochemistry was performed with mouse anti-human CD68 monoclonal antibody at 1:5 dilution (clone KP1; Abcam, Cambridge, UK), rabbit anti-human colony-stimulating factor (CSF)-1 polyclonal antibody at 1:100 dilution (BA0750; Wuhan Boster Biological Technology, Ltd., Wuhan, China), anti-interleukin (IL)-6 at 1:100 dilution (BS0781R; BIOSS, Beijing, China), anti-matrix metalloproteinase (MMP)-2 at 1:400 dilution (ab37150; Abcam), anti-E-cadherin at 1:200 dilution (sc-7,870; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) and anti-Snail at 1:25 dilution (AP2054a; Abgent, Inc., San Diego, CA, USA).

Techniques: Expressing, Staining

Journal: Oncology Letters

Article Title: Significance of interstitial tumor-associated macrophages in the progression of lung adenocarcinoma

doi: 10.3892/ol.2016.5270

Figure Lengend Snippet: (A) Kaplan-Meier analysis of overall survival for the combination of CD68, CSF-1 and IL-6. (B) Kaplan-Meier analysis of disease-free survival for the combination of CD68, CSF-1 and IL-6. CSF-1, colony-stimulating factor-1; CD, cluster of differentiation; IL, interleukin,

Article Snippet: Immunohistochemistry was performed with mouse anti-human CD68 monoclonal antibody at 1:5 dilution (clone KP1; Abcam, Cambridge, UK), rabbit anti-human colony-stimulating factor (CSF)-1 polyclonal antibody at 1:100 dilution (BA0750; Wuhan Boster Biological Technology, Ltd., Wuhan, China), anti-interleukin (IL)-6 at 1:100 dilution (BS0781R; BIOSS, Beijing, China), anti-matrix metalloproteinase (MMP)-2 at 1:400 dilution (ab37150; Abcam), anti-E-cadherin at 1:200 dilution (sc-7,870; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) and anti-Snail at 1:25 dilution (AP2054a; Abgent, Inc., San Diego, CA, USA).

Techniques:

Journal: Oncology Letters

Article Title: Significance of interstitial tumor-associated macrophages in the progression of lung adenocarcinoma

doi: 10.3892/ol.2016.5270

Figure Lengend Snippet: (A) OS in patients with AIS/MIA in the CD68+CSF-1+IL-6+ group compared with other groups, including the CD68-, CD68+CSF-1+IL-6-, CD68+CSF-1-IL-6+ and CD68+CSF-1-IL-6- groups. (B) DFS in patients with AIS/MIA in the CD68+ CSF-1+ IL-6+ group compared with other groups (C) Comparison of OS in AIS/MIA patients with CD68+CSF-1+IL-6+ and in LPA patients with CD68+CSF-1+IL-6+. (D) Comparison of DFS in AIS/MIA patients with CD68+CSF-1+IL-6+ and in LPA patients with CD68+CSF-1+IL-6+. *Others include the CD68+CSF-1+IL-6- group, the CD68+CSF-1-IL-6+ group, the CD68+CSF-1-IL-6- group and the CD68- group. AIS, adenocarcinoma in situ ; MIA, minimally invasive adenocarcinoma; CD, cluster of differentiation; CSF, colony-stimulating factor; IL, interleukin; OS, overall survival; DFS, disease-free survival.

Article Snippet: Immunohistochemistry was performed with mouse anti-human CD68 monoclonal antibody at 1:5 dilution (clone KP1; Abcam, Cambridge, UK), rabbit anti-human colony-stimulating factor (CSF)-1 polyclonal antibody at 1:100 dilution (BA0750; Wuhan Boster Biological Technology, Ltd., Wuhan, China), anti-interleukin (IL)-6 at 1:100 dilution (BS0781R; BIOSS, Beijing, China), anti-matrix metalloproteinase (MMP)-2 at 1:400 dilution (ab37150; Abcam), anti-E-cadherin at 1:200 dilution (sc-7,870; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) and anti-Snail at 1:25 dilution (AP2054a; Abgent, Inc., San Diego, CA, USA).

Techniques: In Situ

Journal: Frontiers in Immunology

Article Title: MSC therapy ameliorates experimental gouty arthritis hinting an early COX-2 induction

doi: 10.3389/fimmu.2023.1193179

Figure Lengend Snippet: Ad-MSC modulates pro- and anti-inflammatory cytokine profile in synovial membranes of arthritic rabbits. Representative western blot of pro-inflammatory cytokines (A) COX-2, TNF, IL-6 and M2 anti-inflammatory cytokines levels (B) IL-10 and TGF-β. EZ blue staining was used as protein loading control and to normalize the results, which are expressed as a fold-change of the Control group. Bars show the mean and SEM. COX-2, Ciclooxygenase-2; IL, interleukin; TGF-β, tumor growth factor-β, TNF-α, tumor necrosis factor α; MSC, mesenchymal stem cells; MSU, monosodium urate.

Article Snippet: The following primary antibodies were applied overnight at 4°C: anti-human COX-2 (Santa Cruz Biotechnology, Dallas TX, USA), anti-rabbit IL-6, anti-rabbit TNF, anti-rabbit IL-10, anti-rabbit TGF-β (all from Cloud-Clone Corp; 1/250 dilution); anti-human NLRP3 (AdipoGen, Liestal, Switzerland; 1/1000 dilution); anti-human Caspase-1 (Thermo Fisher Scientific, IL, USA; 1/500 dilution), anti-rabbit IL-18 and IL-1β antibodies (Cloud-Clone Corp, Houston TX, USA; 1/250 dilution).

Techniques: Western Blot, Staining

Journal: The Journal of Experimental Medicine

Article Title: Assembly and Regulation of the CD40 Receptor Complex in Human B Cells

doi:

Figure Lengend Snippet: Stimulation with sCD154 induces recruitment of TRAF2 and TRAF3 to CD40. ( A ) DND39 cells (2 × 10 7 cells/lane) were either left unstimulated (lanes 1 , 2 , 5 , and 7 ), or stimulated with sCD154 (4 nM) (lanes 3 , 4 , 6 , and 8 ) for 15 min before lysis and immunoprecipitation with either irrelevant mouse IgG1 (lanes 1 , 3 ) and or with anti-human CD40 mouse IgG1 monoclonal BE-1 ( 2 , 4 ) or S2C6 (lanes 5 , 6 , and 7 , 8 ), respectively. The immunoprecipitated samples were immunoblotted for TRAF2 ( arrowhead ) (lanes 1–6 ) or for TRAF3 (indicated by asterisk) (lanes 7 , 8 ). This experiment is representative of 12 experiments.

Article Snippet: Precipitated proteins were separated by SDS-PAGE, transferred to nitrocellulose, and coprecipitated TRAF molecules detected with polyclonal rabbit anti–human TRAF2 (C20) ( Santa Cruz Biotech nologies, Santa Cruz, CA) or anti–human TRAF3 (N) produced against a peptide corresponding to residues 9–32 of the human TRAF3 sequence.

Techniques: Lysis, Immunoprecipitation

Journal: The Journal of Experimental Medicine

Article Title: Assembly and Regulation of the CD40 Receptor Complex in Human B Cells

doi:

Figure Lengend Snippet: Anti-μ cross-linking reduces TRAF2 association and Fas upregulation induced by CD40 signaling. DND39 cells (2 × 10 7 cells/lane) were incubated in media or with 10 μg/ml goat anti–human IgM for 24 h and both groups were either left unstimulated ( minus ) or stimulated with sCD154 (4 nM) ( plus ) for 15 min. Lysates were immunoprecipitated for CD40 and immunoblotted for TRAF2 ( A ) or TRAF3 ( B ). ( C and D ) Immunoblot analysis of cytosolic TRAF content after GST–CD40cyt immunoprecipitations. The lysates used for the anti-CD40 precipitations described in A and B were further precipitated with GST–CD40cyt. 5 × 10 6 cells/lane were loaded for the TRAF2 blot, while the TRAF3 blot received 2 × 10 7 cell equivalents per lane. ( E ) Fluorescence-activated cell sorting analysis of DND39 cells after treatment with anti-IgM and sCD154. DND39 cells were incubated for 24 h with media ( minus ), 10 μg/ml anti-IgM, sCD154, or both (as indicated). Fas expression was detected with anti-CD95 monoclonal antibody ( open profile ) or a control mouse IgG1 monoclonal ( closed profile ). The mean fluorescent intensity of the cells is indicated in the upper righthand corner. This experiment is representative of four experiments.

Article Snippet: Precipitated proteins were separated by SDS-PAGE, transferred to nitrocellulose, and coprecipitated TRAF molecules detected with polyclonal rabbit anti–human TRAF2 (C20) ( Santa Cruz Biotech nologies, Santa Cruz, CA) or anti–human TRAF3 (N) produced against a peptide corresponding to residues 9–32 of the human TRAF3 sequence.

Techniques: Incubation, Immunoprecipitation, Western Blot, Fluorescence, FACS, Expressing, Control

Journal: The Journal of Experimental Medicine

Article Title: Assembly and Regulation of the CD40 Receptor Complex in Human B Cells

doi:

Figure Lengend Snippet: Engagement of CD40 results in the loss of immunoprecipitable TRAF2 and TRAF3 from DND39 cells. DND39 cells (2 × 10 6 cells/ml) were left untreated ( minus ) or treated with 4 nM sCD154 ( plus ) for 15 min. After treatment, lysates were immunoprecipitated with either anti-TRAF2 or anti-TRAF3 antibodies. 5 × 10 6 cell equivalents were loaded per lane.

Article Snippet: Precipitated proteins were separated by SDS-PAGE, transferred to nitrocellulose, and coprecipitated TRAF molecules detected with polyclonal rabbit anti–human TRAF2 (C20) ( Santa Cruz Biotech nologies, Santa Cruz, CA) or anti–human TRAF3 (N) produced against a peptide corresponding to residues 9–32 of the human TRAF3 sequence.

Techniques: Immunoprecipitation

Journal: The Journal of Experimental Medicine

Article Title: Assembly and Regulation of the CD40 Receptor Complex in Human B Cells

doi:

Figure Lengend Snippet: Pretreatment with IL-4 increases TRAF2, but not TRAF3, recruited to CD40 and increases cell surface Fas expression. DND39 cells (2 × 10 7 cells/lane) were either left untreated or pretreated for 10 min with human (h)IL-4 (2 ng/ml) (Genzyme, Cambridge, MA); and both groups were either left unstimulated ( minus ) or stimulated with sCD154 (4 nM) ( plus ) for 15 min. Lysates were immunoprecipitated for CD40 and immunoblotted for TRAF2 ( A ) or TRAF3 ( B ). ( C ) Fluorescence-activated cell sorting analysis of DND39 cells after treatment with hIL-4 and sCD154. DND39 cells were incubated for 24 h with media ( minus ), 2 ng/ml hIL-4, sCD154, or both (as indicated). Fas expression was detected with anti-CD95 monoclonal antibody ( open profile ) or a control mouse IgG1 monoclonal ( closed profile ). The mean fluorescent intensity of the cells is indicated in the upper righthand corner. This experiment is representative of four experiments.

Article Snippet: Precipitated proteins were separated by SDS-PAGE, transferred to nitrocellulose, and coprecipitated TRAF molecules detected with polyclonal rabbit anti–human TRAF2 (C20) ( Santa Cruz Biotech nologies, Santa Cruz, CA) or anti–human TRAF3 (N) produced against a peptide corresponding to residues 9–32 of the human TRAF3 sequence.

Techniques: Expressing, Immunoprecipitation, Fluorescence, FACS, Incubation, Control