mouse cd14 immunoassay kit Search Results


cd14 microbeads  (Miltenyi Biotec)
98
Miltenyi Biotec cd14 microbeads
Human SI contains four distinct Mf subsets. (A) Representative flow cytometric analysis of human SI Mfs, identifying Mf1 (purple) and Mf2 (green, top right), Mf3 (red), and Mf4 (blue, bottom right), and DCs (pink, top center) among CD45 + HLA-DR + SI mononuclear phagocytes. Arrows indicate sequential gating (see Fig. S1 A for extended gating strategy). Dot plots are representative of all subjects. (B) The proportion of SI mononuclear phagocyte subsets given as percentage of all Mfs and DCs (CD45 + HLA-DR + ) was determined by flow cytometry; bars indicate median values ( n = 55). (C) Representative micrographs of SI Mf subsets sorted by flow cytometry and stained with Hemacolor reagent. n = 2. Bars, 5 µm. (D) Representative dot plots showing FSC-A (size), SSC-A (granularity), and autofluorescence of SI Mf subsets and DCs. Autofluorescence was measured as fluorescence excited by a 488-nm laser and collected by a 530/30 bandpass filter, where matched fluorochrome was not included. Data are representative of all subjects. (E) Representative flow cytometric staining of surface markers on PBDC, <t>CD14</t> + PBMo ( n = 3 or more; gated as in Fig. S1 B), and SI Mf subsets and DCs ( n = 4 or more). Gray histograms represent FMO controls.
Cd14 Microbeads, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+cd14+immunoassay+kit/pmc05789404-221-35-37?v=Miltenyi+Biotec
Average 98 stars, based on 1 article reviews
cd14 microbeads - by Bioz Stars, 2026-08
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mouse cd14 immunoassay  (R&D Systems)
94
R&D Systems mouse cd14 immunoassay
A Representative experiment of 3 independent experiments of IAP expression levels analyzed by Western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. B Quantification of GL261-DsRed spheroids area upon vehicle ( n = 10) and SMg treatment ( n = 9) and in the presence or not of the C8B4 cells ( n = 10). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. C Concentrations of <t>CD14</t> and CCL17/TARC quantified in the supernatant of C8B4 and GL261-DsRed co-cultures by ELISA assay after 72 h of vehicle or SMg treatments ( n = 6 independent experiments). Statistical analyses were performed by using Mann–Whitney test; alpha = 0.05, bilateral p-value: ** p < 0.005. D Quantification of GL261-DsRed spheroids area in the presence of the C8B4 cells expressing ML-IAP (siCTRL) or down-expressing ML-IAP (siML-IAP) ( n = 10) after 72 h of treatment vehicle ( n = 10) and SMg treatment ( n = 9). Data from three independent experiments. Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: * p < 0.05 ; ** p < 0.005 ; **** p < 0.0001. E Quantification of GL261-DsRed spheroid area in the presence of C8B4 cells treated with vehicle and SMg for 72 h. C8B4 cells were pre-treated for 24 h with ZVAD (vehicle n = 23, SMg n = 23) and TNFαi (vehicle n = 27, SMg n = 23) or without pre-treatment (vehicle n = 28, SMg n = 25). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. F Representative experiment ( n = 2) of IAP expression levels analyzed by western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. G Representative experiment ( n = 2) of expression levels of p65, phospho-p65, IκBα, phospho IκB, caspase-3, cleaved caspase-3 analyzed by Western blotting in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. H Representative experiment out of 3 experiments of expression levels of CD206 and iNOS analyzed by Western blotting after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Expression level of actin β served as loading control. I iNOS/CD206 ratio in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Quantification was performed from 3 independent experiments using ImageJ software and data presented were normalized to actin β expression. iNOS/CD206 ratio fold changes were normalized on vehicle condition. B – E , I Bar graphs represent mean ± s.e.m.
Mouse Cd14 Immunoassay, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 94 stars, based on 1 article reviews
mouse cd14 immunoassay - by Bioz Stars, 2026-08
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quantikine elisa human il 22 immunoassay  (R&D Systems)
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R&D Systems quantikine elisa human il 22 immunoassay
A Representative experiment of 3 independent experiments of IAP expression levels analyzed by Western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. B Quantification of GL261-DsRed spheroids area upon vehicle ( n = 10) and SMg treatment ( n = 9) and in the presence or not of the C8B4 cells ( n = 10). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. C Concentrations of <t>CD14</t> and CCL17/TARC quantified in the supernatant of C8B4 and GL261-DsRed co-cultures by ELISA assay after 72 h of vehicle or SMg treatments ( n = 6 independent experiments). Statistical analyses were performed by using Mann–Whitney test; alpha = 0.05, bilateral p-value: ** p < 0.005. D Quantification of GL261-DsRed spheroids area in the presence of the C8B4 cells expressing ML-IAP (siCTRL) or down-expressing ML-IAP (siML-IAP) ( n = 10) after 72 h of treatment vehicle ( n = 10) and SMg treatment ( n = 9). Data from three independent experiments. Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: * p < 0.05 ; ** p < 0.005 ; **** p < 0.0001. E Quantification of GL261-DsRed spheroid area in the presence of C8B4 cells treated with vehicle and SMg for 72 h. C8B4 cells were pre-treated for 24 h with ZVAD (vehicle n = 23, SMg n = 23) and TNFαi (vehicle n = 27, SMg n = 23) or without pre-treatment (vehicle n = 28, SMg n = 25). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. F Representative experiment ( n = 2) of IAP expression levels analyzed by western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. G Representative experiment ( n = 2) of expression levels of p65, phospho-p65, IκBα, phospho IκB, caspase-3, cleaved caspase-3 analyzed by Western blotting in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. H Representative experiment out of 3 experiments of expression levels of CD206 and iNOS analyzed by Western blotting after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Expression level of actin β served as loading control. I iNOS/CD206 ratio in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Quantification was performed from 3 independent experiments using ImageJ software and data presented were normalized to actin β expression. iNOS/CD206 ratio fold changes were normalized on vehicle condition. B – E , I Bar graphs represent mean ± s.e.m.
Quantikine Elisa Human Il 22 Immunoassay, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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quantikine m mouse immunoassays kits  (R&D Systems)
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R&D Systems quantikine m mouse immunoassays kits
A Representative experiment of 3 independent experiments of IAP expression levels analyzed by Western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. B Quantification of GL261-DsRed spheroids area upon vehicle ( n = 10) and SMg treatment ( n = 9) and in the presence or not of the C8B4 cells ( n = 10). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. C Concentrations of <t>CD14</t> and CCL17/TARC quantified in the supernatant of C8B4 and GL261-DsRed co-cultures by ELISA assay after 72 h of vehicle or SMg treatments ( n = 6 independent experiments). Statistical analyses were performed by using Mann–Whitney test; alpha = 0.05, bilateral p-value: ** p < 0.005. D Quantification of GL261-DsRed spheroids area in the presence of the C8B4 cells expressing ML-IAP (siCTRL) or down-expressing ML-IAP (siML-IAP) ( n = 10) after 72 h of treatment vehicle ( n = 10) and SMg treatment ( n = 9). Data from three independent experiments. Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: * p < 0.05 ; ** p < 0.005 ; **** p < 0.0001. E Quantification of GL261-DsRed spheroid area in the presence of C8B4 cells treated with vehicle and SMg for 72 h. C8B4 cells were pre-treated for 24 h with ZVAD (vehicle n = 23, SMg n = 23) and TNFαi (vehicle n = 27, SMg n = 23) or without pre-treatment (vehicle n = 28, SMg n = 25). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. F Representative experiment ( n = 2) of IAP expression levels analyzed by western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. G Representative experiment ( n = 2) of expression levels of p65, phospho-p65, IκBα, phospho IκB, caspase-3, cleaved caspase-3 analyzed by Western blotting in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. H Representative experiment out of 3 experiments of expression levels of CD206 and iNOS analyzed by Western blotting after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Expression level of actin β served as loading control. I iNOS/CD206 ratio in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Quantification was performed from 3 independent experiments using ImageJ software and data presented were normalized to actin β expression. iNOS/CD206 ratio fold changes were normalized on vehicle condition. B – E , I Bar graphs represent mean ± s.e.m.
Quantikine M Mouse Immunoassays Kits, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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quantikine m mouse immunoassays kits - by Bioz Stars, 2026-08
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mouse ccl17/tarc quantikine elisa kit  (Bio-Techne corporation)
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Bio-Techne corporation mouse ccl17/tarc quantikine elisa kit
A Representative experiment of 3 independent experiments of IAP expression levels analyzed by Western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. B Quantification of GL261-DsRed spheroids area upon vehicle ( n = 10) and SMg treatment ( n = 9) and in the presence or not of the C8B4 cells ( n = 10). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. C Concentrations of <t>CD14</t> and CCL17/TARC quantified in the supernatant of C8B4 and GL261-DsRed co-cultures by ELISA assay after 72 h of vehicle or SMg treatments ( n = 6 independent experiments). Statistical analyses were performed by using Mann–Whitney test; alpha = 0.05, bilateral p-value: ** p < 0.005. D Quantification of GL261-DsRed spheroids area in the presence of the C8B4 cells expressing ML-IAP (siCTRL) or down-expressing ML-IAP (siML-IAP) ( n = 10) after 72 h of treatment vehicle ( n = 10) and SMg treatment ( n = 9). Data from three independent experiments. Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: * p < 0.05 ; ** p < 0.005 ; **** p < 0.0001. E Quantification of GL261-DsRed spheroid area in the presence of C8B4 cells treated with vehicle and SMg for 72 h. C8B4 cells were pre-treated for 24 h with ZVAD (vehicle n = 23, SMg n = 23) and TNFαi (vehicle n = 27, SMg n = 23) or without pre-treatment (vehicle n = 28, SMg n = 25). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. F Representative experiment ( n = 2) of IAP expression levels analyzed by western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. G Representative experiment ( n = 2) of expression levels of p65, phospho-p65, IκBα, phospho IκB, caspase-3, cleaved caspase-3 analyzed by Western blotting in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. H Representative experiment out of 3 experiments of expression levels of CD206 and iNOS analyzed by Western blotting after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Expression level of actin β served as loading control. I iNOS/CD206 ratio in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Quantification was performed from 3 independent experiments using ImageJ software and data presented were normalized to actin β expression. iNOS/CD206 ratio fold changes were normalized on vehicle condition. B – E , I Bar graphs represent mean ± s.e.m.
Mouse Ccl17/Tarc Quantikine Elisa Kit, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse ccl17/tarc quantikine elisa kit - by Bioz Stars, 2026-08
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human cxcl12/sdf-1 alpha quantikine elisa kit  (Bio-Techne corporation)
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Bio-Techne corporation human cxcl12/sdf-1 alpha quantikine elisa kit
A Representative experiment of 3 independent experiments of IAP expression levels analyzed by Western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. B Quantification of GL261-DsRed spheroids area upon vehicle ( n = 10) and SMg treatment ( n = 9) and in the presence or not of the C8B4 cells ( n = 10). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. C Concentrations of <t>CD14</t> and CCL17/TARC quantified in the supernatant of C8B4 and GL261-DsRed co-cultures by ELISA assay after 72 h of vehicle or SMg treatments ( n = 6 independent experiments). Statistical analyses were performed by using Mann–Whitney test; alpha = 0.05, bilateral p-value: ** p < 0.005. D Quantification of GL261-DsRed spheroids area in the presence of the C8B4 cells expressing ML-IAP (siCTRL) or down-expressing ML-IAP (siML-IAP) ( n = 10) after 72 h of treatment vehicle ( n = 10) and SMg treatment ( n = 9). Data from three independent experiments. Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: * p < 0.05 ; ** p < 0.005 ; **** p < 0.0001. E Quantification of GL261-DsRed spheroid area in the presence of C8B4 cells treated with vehicle and SMg for 72 h. C8B4 cells were pre-treated for 24 h with ZVAD (vehicle n = 23, SMg n = 23) and TNFαi (vehicle n = 27, SMg n = 23) or without pre-treatment (vehicle n = 28, SMg n = 25). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. F Representative experiment ( n = 2) of IAP expression levels analyzed by western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. G Representative experiment ( n = 2) of expression levels of p65, phospho-p65, IκBα, phospho IκB, caspase-3, cleaved caspase-3 analyzed by Western blotting in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. H Representative experiment out of 3 experiments of expression levels of CD206 and iNOS analyzed by Western blotting after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Expression level of actin β served as loading control. I iNOS/CD206 ratio in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Quantification was performed from 3 independent experiments using ImageJ software and data presented were normalized to actin β expression. iNOS/CD206 ratio fold changes were normalized on vehicle condition. B – E , I Bar graphs represent mean ± s.e.m.
Human Cxcl12/Sdf 1 Alpha Quantikine Elisa Kit, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti-human cd14 antibody  (Unipath Limited)
90
Unipath Limited anti-human cd14 antibody
A Representative experiment of 3 independent experiments of IAP expression levels analyzed by Western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. B Quantification of GL261-DsRed spheroids area upon vehicle ( n = 10) and SMg treatment ( n = 9) and in the presence or not of the C8B4 cells ( n = 10). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. C Concentrations of <t>CD14</t> and CCL17/TARC quantified in the supernatant of C8B4 and GL261-DsRed co-cultures by ELISA assay after 72 h of vehicle or SMg treatments ( n = 6 independent experiments). Statistical analyses were performed by using Mann–Whitney test; alpha = 0.05, bilateral p-value: ** p < 0.005. D Quantification of GL261-DsRed spheroids area in the presence of the C8B4 cells expressing ML-IAP (siCTRL) or down-expressing ML-IAP (siML-IAP) ( n = 10) after 72 h of treatment vehicle ( n = 10) and SMg treatment ( n = 9). Data from three independent experiments. Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: * p < 0.05 ; ** p < 0.005 ; **** p < 0.0001. E Quantification of GL261-DsRed spheroid area in the presence of C8B4 cells treated with vehicle and SMg for 72 h. C8B4 cells were pre-treated for 24 h with ZVAD (vehicle n = 23, SMg n = 23) and TNFαi (vehicle n = 27, SMg n = 23) or without pre-treatment (vehicle n = 28, SMg n = 25). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. F Representative experiment ( n = 2) of IAP expression levels analyzed by western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. G Representative experiment ( n = 2) of expression levels of p65, phospho-p65, IκBα, phospho IκB, caspase-3, cleaved caspase-3 analyzed by Western blotting in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. H Representative experiment out of 3 experiments of expression levels of CD206 and iNOS analyzed by Western blotting after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Expression level of actin β served as loading control. I iNOS/CD206 ratio in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Quantification was performed from 3 independent experiments using ImageJ software and data presented were normalized to actin β expression. iNOS/CD206 ratio fold changes were normalized on vehicle condition. B – E , I Bar graphs represent mean ± s.e.m.
Anti Human Cd14 Antibody, supplied by Unipath Limited, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+cd14+immunoassay+kit/10__1182_slash_blood__v91__7__2565-41-32-36?v=Unipath+Limited
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anti-human cd14 antibody - by Bioz Stars, 2026-08
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fetal calf serum (fcs)  (ICN Biomedicals)
90
ICN Biomedicals fetal calf serum (fcs)
A Representative experiment of 3 independent experiments of IAP expression levels analyzed by Western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. B Quantification of GL261-DsRed spheroids area upon vehicle ( n = 10) and SMg treatment ( n = 9) and in the presence or not of the C8B4 cells ( n = 10). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. C Concentrations of <t>CD14</t> and CCL17/TARC quantified in the supernatant of C8B4 and GL261-DsRed co-cultures by ELISA assay after 72 h of vehicle or SMg treatments ( n = 6 independent experiments). Statistical analyses were performed by using Mann–Whitney test; alpha = 0.05, bilateral p-value: ** p < 0.005. D Quantification of GL261-DsRed spheroids area in the presence of the C8B4 cells expressing ML-IAP (siCTRL) or down-expressing ML-IAP (siML-IAP) ( n = 10) after 72 h of treatment vehicle ( n = 10) and SMg treatment ( n = 9). Data from three independent experiments. Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: * p < 0.05 ; ** p < 0.005 ; **** p < 0.0001. E Quantification of GL261-DsRed spheroid area in the presence of C8B4 cells treated with vehicle and SMg for 72 h. C8B4 cells were pre-treated for 24 h with ZVAD (vehicle n = 23, SMg n = 23) and TNFαi (vehicle n = 27, SMg n = 23) or without pre-treatment (vehicle n = 28, SMg n = 25). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. F Representative experiment ( n = 2) of IAP expression levels analyzed by western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. G Representative experiment ( n = 2) of expression levels of p65, phospho-p65, IκBα, phospho IκB, caspase-3, cleaved caspase-3 analyzed by Western blotting in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. H Representative experiment out of 3 experiments of expression levels of CD206 and iNOS analyzed by Western blotting after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Expression level of actin β served as loading control. I iNOS/CD206 ratio in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Quantification was performed from 3 independent experiments using ImageJ software and data presented were normalized to actin β expression. iNOS/CD206 ratio fold changes were normalized on vehicle condition. B – E , I Bar graphs represent mean ± s.e.m.
Fetal Calf Serum (Fcs), supplied by ICN Biomedicals, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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facsaria fusion flow cytometer  (Becton Dickinson)
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Becton Dickinson facsaria fusion flow cytometer
A Representative experiment of 3 independent experiments of IAP expression levels analyzed by Western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. B Quantification of GL261-DsRed spheroids area upon vehicle ( n = 10) and SMg treatment ( n = 9) and in the presence or not of the C8B4 cells ( n = 10). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. C Concentrations of <t>CD14</t> and CCL17/TARC quantified in the supernatant of C8B4 and GL261-DsRed co-cultures by ELISA assay after 72 h of vehicle or SMg treatments ( n = 6 independent experiments). Statistical analyses were performed by using Mann–Whitney test; alpha = 0.05, bilateral p-value: ** p < 0.005. D Quantification of GL261-DsRed spheroids area in the presence of the C8B4 cells expressing ML-IAP (siCTRL) or down-expressing ML-IAP (siML-IAP) ( n = 10) after 72 h of treatment vehicle ( n = 10) and SMg treatment ( n = 9). Data from three independent experiments. Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: * p < 0.05 ; ** p < 0.005 ; **** p < 0.0001. E Quantification of GL261-DsRed spheroid area in the presence of C8B4 cells treated with vehicle and SMg for 72 h. C8B4 cells were pre-treated for 24 h with ZVAD (vehicle n = 23, SMg n = 23) and TNFαi (vehicle n = 27, SMg n = 23) or without pre-treatment (vehicle n = 28, SMg n = 25). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. F Representative experiment ( n = 2) of IAP expression levels analyzed by western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. G Representative experiment ( n = 2) of expression levels of p65, phospho-p65, IκBα, phospho IκB, caspase-3, cleaved caspase-3 analyzed by Western blotting in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. H Representative experiment out of 3 experiments of expression levels of CD206 and iNOS analyzed by Western blotting after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Expression level of actin β served as loading control. I iNOS/CD206 ratio in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Quantification was performed from 3 independent experiments using ImageJ software and data presented were normalized to actin β expression. iNOS/CD206 ratio fold changes were normalized on vehicle condition. B – E , I Bar graphs represent mean ± s.e.m.
Facsaria Fusion Flow Cytometer, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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magic-fer radioimmunoassay kit for ferritin  (Corning Life Sciences)
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A Representative experiment of 3 independent experiments of IAP expression levels analyzed by Western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. B Quantification of GL261-DsRed spheroids area upon vehicle ( n = 10) and SMg treatment ( n = 9) and in the presence or not of the C8B4 cells ( n = 10). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. C Concentrations of <t>CD14</t> and CCL17/TARC quantified in the supernatant of C8B4 and GL261-DsRed co-cultures by ELISA assay after 72 h of vehicle or SMg treatments ( n = 6 independent experiments). Statistical analyses were performed by using Mann–Whitney test; alpha = 0.05, bilateral p-value: ** p < 0.005. D Quantification of GL261-DsRed spheroids area in the presence of the C8B4 cells expressing ML-IAP (siCTRL) or down-expressing ML-IAP (siML-IAP) ( n = 10) after 72 h of treatment vehicle ( n = 10) and SMg treatment ( n = 9). Data from three independent experiments. Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: * p < 0.05 ; ** p < 0.005 ; **** p < 0.0001. E Quantification of GL261-DsRed spheroid area in the presence of C8B4 cells treated with vehicle and SMg for 72 h. C8B4 cells were pre-treated for 24 h with ZVAD (vehicle n = 23, SMg n = 23) and TNFαi (vehicle n = 27, SMg n = 23) or without pre-treatment (vehicle n = 28, SMg n = 25). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. F Representative experiment ( n = 2) of IAP expression levels analyzed by western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. G Representative experiment ( n = 2) of expression levels of p65, phospho-p65, IκBα, phospho IκB, caspase-3, cleaved caspase-3 analyzed by Western blotting in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. H Representative experiment out of 3 experiments of expression levels of CD206 and iNOS analyzed by Western blotting after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Expression level of actin β served as loading control. I iNOS/CD206 ratio in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Quantification was performed from 3 independent experiments using ImageJ software and data presented were normalized to actin β expression. iNOS/CD206 ratio fold changes were normalized on vehicle condition. B – E , I Bar graphs represent mean ± s.e.m.
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A Representative experiment of 3 independent experiments of IAP expression levels analyzed by Western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. B Quantification of GL261-DsRed spheroids area upon vehicle ( n = 10) and SMg treatment ( n = 9) and in the presence or not of the C8B4 cells ( n = 10). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. C Concentrations of <t>CD14</t> and CCL17/TARC quantified in the supernatant of C8B4 and GL261-DsRed co-cultures by ELISA assay after 72 h of vehicle or SMg treatments ( n = 6 independent experiments). Statistical analyses were performed by using Mann–Whitney test; alpha = 0.05, bilateral p-value: ** p < 0.005. D Quantification of GL261-DsRed spheroids area in the presence of the C8B4 cells expressing ML-IAP (siCTRL) or down-expressing ML-IAP (siML-IAP) ( n = 10) after 72 h of treatment vehicle ( n = 10) and SMg treatment ( n = 9). Data from three independent experiments. Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: * p < 0.05 ; ** p < 0.005 ; **** p < 0.0001. E Quantification of GL261-DsRed spheroid area in the presence of C8B4 cells treated with vehicle and SMg for 72 h. C8B4 cells were pre-treated for 24 h with ZVAD (vehicle n = 23, SMg n = 23) and TNFαi (vehicle n = 27, SMg n = 23) or without pre-treatment (vehicle n = 28, SMg n = 25). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. F Representative experiment ( n = 2) of IAP expression levels analyzed by western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. G Representative experiment ( n = 2) of expression levels of p65, phospho-p65, IκBα, phospho IκB, caspase-3, cleaved caspase-3 analyzed by Western blotting in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. H Representative experiment out of 3 experiments of expression levels of CD206 and iNOS analyzed by Western blotting after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Expression level of actin β served as loading control. I iNOS/CD206 ratio in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Quantification was performed from 3 independent experiments using ImageJ software and data presented were normalized to actin β expression. iNOS/CD206 ratio fold changes were normalized on vehicle condition. B – E , I Bar graphs represent mean ± s.e.m.
Rpmi 1640, supplied by ICN Biomedicals, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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A Representative experiment of 3 independent experiments of IAP expression levels analyzed by Western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. B Quantification of GL261-DsRed spheroids area upon vehicle ( n = 10) and SMg treatment ( n = 9) and in the presence or not of the C8B4 cells ( n = 10). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. C Concentrations of <t>CD14</t> and CCL17/TARC quantified in the supernatant of C8B4 and GL261-DsRed co-cultures by ELISA assay after 72 h of vehicle or SMg treatments ( n = 6 independent experiments). Statistical analyses were performed by using Mann–Whitney test; alpha = 0.05, bilateral p-value: ** p < 0.005. D Quantification of GL261-DsRed spheroids area in the presence of the C8B4 cells expressing ML-IAP (siCTRL) or down-expressing ML-IAP (siML-IAP) ( n = 10) after 72 h of treatment vehicle ( n = 10) and SMg treatment ( n = 9). Data from three independent experiments. Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: * p < 0.05 ; ** p < 0.005 ; **** p < 0.0001. E Quantification of GL261-DsRed spheroid area in the presence of C8B4 cells treated with vehicle and SMg for 72 h. C8B4 cells were pre-treated for 24 h with ZVAD (vehicle n = 23, SMg n = 23) and TNFαi (vehicle n = 27, SMg n = 23) or without pre-treatment (vehicle n = 28, SMg n = 25). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. F Representative experiment ( n = 2) of IAP expression levels analyzed by western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. G Representative experiment ( n = 2) of expression levels of p65, phospho-p65, IκBα, phospho IκB, caspase-3, cleaved caspase-3 analyzed by Western blotting in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. H Representative experiment out of 3 experiments of expression levels of CD206 and iNOS analyzed by Western blotting after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Expression level of actin β served as loading control. I iNOS/CD206 ratio in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Quantification was performed from 3 independent experiments using ImageJ software and data presented were normalized to actin β expression. iNOS/CD206 ratio fold changes were normalized on vehicle condition. B – E , I Bar graphs represent mean ± s.e.m.
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Image Search Results


Human SI contains four distinct Mf subsets. (A) Representative flow cytometric analysis of human SI Mfs, identifying Mf1 (purple) and Mf2 (green, top right), Mf3 (red), and Mf4 (blue, bottom right), and DCs (pink, top center) among CD45 + HLA-DR + SI mononuclear phagocytes. Arrows indicate sequential gating (see Fig. S1 A for extended gating strategy). Dot plots are representative of all subjects. (B) The proportion of SI mononuclear phagocyte subsets given as percentage of all Mfs and DCs (CD45 + HLA-DR + ) was determined by flow cytometry; bars indicate median values ( n = 55). (C) Representative micrographs of SI Mf subsets sorted by flow cytometry and stained with Hemacolor reagent. n = 2. Bars, 5 µm. (D) Representative dot plots showing FSC-A (size), SSC-A (granularity), and autofluorescence of SI Mf subsets and DCs. Autofluorescence was measured as fluorescence excited by a 488-nm laser and collected by a 530/30 bandpass filter, where matched fluorochrome was not included. Data are representative of all subjects. (E) Representative flow cytometric staining of surface markers on PBDC, CD14 + PBMo ( n = 3 or more; gated as in Fig. S1 B), and SI Mf subsets and DCs ( n = 4 or more). Gray histograms represent FMO controls.

Journal: The Journal of Experimental Medicine

Article Title: Transcriptional and functional profiling defines human small intestinal macrophage subsets

doi: 10.1084/jem.20170057

Figure Lengend Snippet: Human SI contains four distinct Mf subsets. (A) Representative flow cytometric analysis of human SI Mfs, identifying Mf1 (purple) and Mf2 (green, top right), Mf3 (red), and Mf4 (blue, bottom right), and DCs (pink, top center) among CD45 + HLA-DR + SI mononuclear phagocytes. Arrows indicate sequential gating (see Fig. S1 A for extended gating strategy). Dot plots are representative of all subjects. (B) The proportion of SI mononuclear phagocyte subsets given as percentage of all Mfs and DCs (CD45 + HLA-DR + ) was determined by flow cytometry; bars indicate median values ( n = 55). (C) Representative micrographs of SI Mf subsets sorted by flow cytometry and stained with Hemacolor reagent. n = 2. Bars, 5 µm. (D) Representative dot plots showing FSC-A (size), SSC-A (granularity), and autofluorescence of SI Mf subsets and DCs. Autofluorescence was measured as fluorescence excited by a 488-nm laser and collected by a 530/30 bandpass filter, where matched fluorochrome was not included. Data are representative of all subjects. (E) Representative flow cytometric staining of surface markers on PBDC, CD14 + PBMo ( n = 3 or more; gated as in Fig. S1 B), and SI Mf subsets and DCs ( n = 4 or more). Gray histograms represent FMO controls.

Article Snippet: For cytokine immunoassay and generation of CFSE-monocytes, CD14 + PBMos were sorted from PBMCs (where specified after incubation with Liberase TL and DNase I as described in the Tissue handling and cell isolation section) with CD14 MicroBeads (Miltenyi Biotec).

Techniques: Flow Cytometry, Staining, Fluorescence

Human SI Mf compartment comprises short- and long-lived subsets that differ in marker expression. (A) The percentage of CCR2 + cells among CD14 + PBMo ( n = 5) and SI Mf subsets ( n = 10) was determined by flow cytometry (left). Gates were set according to FMO controls. The expression of CCR2 mRNA was determined by RNaseq analysis; n = 5 (right). (B) The percentage of calprotectin + cells among CD14 + PBMo ( n = 7) and SI Mf subsets ( n = 11) was determined by intracellular staining and flow cytometry. Gates were set according to staining with isotype control antibody. The expression levels of S100A8 and S100A9 mRNA were determined by RNaseq analysis; n = 5 (right). (C) The median fluorescence intensity (MFI) of CD209 relative to MFI of the FMO control on CD14 + PBMo ( n = 5) and SI Mf subsets ( n = 9) was determined by flow cytometry. The expression of CD209 mRNA was determined by RNaseq analysis; n = 5 (right). (A–C, left) Bars represent median values; ns, not significant ( t test comparing CD14 + PBMo and Mf1). *, P < 0.05; ***, P < 0.001; ****, P < 0.0001 (RM-ANOVA of SI subsets). (A–C, right) Expression values (log2FPKM) were mean-centered by transcript. (D) Representative flow cytometric analysis of recipient-specific HLA-A2 staining on Mf subsets from transplanted duodenum 6 wk after transplantation. Numbers on plots represent recipient + cells (mean ± SD from all patients analyzed at this time point; n = 5). Gray histograms represent staining of HLA-DR + stromal cells from the same sample. (E) The percentage of recipient-derived cells in each Mf subset at 3 ( n = 6), 6 ( n = 5), and 52 wk ( n = 6) after transplantation was determined by flow cytometry. Bars indicate median values. *, P < 0.05; ***, P < 0.001; ****, P < 0.0001 (two-way ANOVA with RM on population).

Journal: The Journal of Experimental Medicine

Article Title: Transcriptional and functional profiling defines human small intestinal macrophage subsets

doi: 10.1084/jem.20170057

Figure Lengend Snippet: Human SI Mf compartment comprises short- and long-lived subsets that differ in marker expression. (A) The percentage of CCR2 + cells among CD14 + PBMo ( n = 5) and SI Mf subsets ( n = 10) was determined by flow cytometry (left). Gates were set according to FMO controls. The expression of CCR2 mRNA was determined by RNaseq analysis; n = 5 (right). (B) The percentage of calprotectin + cells among CD14 + PBMo ( n = 7) and SI Mf subsets ( n = 11) was determined by intracellular staining and flow cytometry. Gates were set according to staining with isotype control antibody. The expression levels of S100A8 and S100A9 mRNA were determined by RNaseq analysis; n = 5 (right). (C) The median fluorescence intensity (MFI) of CD209 relative to MFI of the FMO control on CD14 + PBMo ( n = 5) and SI Mf subsets ( n = 9) was determined by flow cytometry. The expression of CD209 mRNA was determined by RNaseq analysis; n = 5 (right). (A–C, left) Bars represent median values; ns, not significant ( t test comparing CD14 + PBMo and Mf1). *, P < 0.05; ***, P < 0.001; ****, P < 0.0001 (RM-ANOVA of SI subsets). (A–C, right) Expression values (log2FPKM) were mean-centered by transcript. (D) Representative flow cytometric analysis of recipient-specific HLA-A2 staining on Mf subsets from transplanted duodenum 6 wk after transplantation. Numbers on plots represent recipient + cells (mean ± SD from all patients analyzed at this time point; n = 5). Gray histograms represent staining of HLA-DR + stromal cells from the same sample. (E) The percentage of recipient-derived cells in each Mf subset at 3 ( n = 6), 6 ( n = 5), and 52 wk ( n = 6) after transplantation was determined by flow cytometry. Bars indicate median values. *, P < 0.05; ***, P < 0.001; ****, P < 0.0001 (two-way ANOVA with RM on population).

Article Snippet: For cytokine immunoassay and generation of CFSE-monocytes, CD14 + PBMos were sorted from PBMCs (where specified after incubation with Liberase TL and DNase I as described in the Tissue handling and cell isolation section) with CD14 MicroBeads (Miltenyi Biotec).

Techniques: Marker, Expressing, Flow Cytometry, Staining, Control, Fluorescence, Transplantation Assay, Derivative Assay

SI Mf subsets are distinguished by their gene expression patterns. (A) PCA of top 1,000 differentially expressed genes within CD14 + PBMo, PBMo-lib (enzyme-treated CD14 + PBMos), SI Mf, and DC subsets and in vitro differentiated Mfs. (B) Pearson correlation and hierarchical clustering analysis of CD14 + PBMo, PBMo-lib, SI Mf, and DC subsets and in vitro Mfs. Fig. S4 D lists all correlation values. (C) Polytomous analysis of differentially expressed genes among CD14 + PBMo and SI Mf and DC subsets, grouped into 16 clusters based on top models obtained from unsupervised clustering. Genes were assigned to a model based on the highest calculated posterior probability value (>0.65). Expression values are log2FPKM. Top-ranked genes from each cluster are listed on the right (full list available in Table S1). (A–C) CD14 + PBMos and SI Mfs were isolated from five donors, SI DC subsets from two of these. PBMo-lib were derived from two of the included CD14 + PBMo samples. In vitro Mfs were differentiated from PBMos of two separate healthy donors.

Journal: The Journal of Experimental Medicine

Article Title: Transcriptional and functional profiling defines human small intestinal macrophage subsets

doi: 10.1084/jem.20170057

Figure Lengend Snippet: SI Mf subsets are distinguished by their gene expression patterns. (A) PCA of top 1,000 differentially expressed genes within CD14 + PBMo, PBMo-lib (enzyme-treated CD14 + PBMos), SI Mf, and DC subsets and in vitro differentiated Mfs. (B) Pearson correlation and hierarchical clustering analysis of CD14 + PBMo, PBMo-lib, SI Mf, and DC subsets and in vitro Mfs. Fig. S4 D lists all correlation values. (C) Polytomous analysis of differentially expressed genes among CD14 + PBMo and SI Mf and DC subsets, grouped into 16 clusters based on top models obtained from unsupervised clustering. Genes were assigned to a model based on the highest calculated posterior probability value (>0.65). Expression values are log2FPKM. Top-ranked genes from each cluster are listed on the right (full list available in Table S1). (A–C) CD14 + PBMos and SI Mfs were isolated from five donors, SI DC subsets from two of these. PBMo-lib were derived from two of the included CD14 + PBMo samples. In vitro Mfs were differentiated from PBMos of two separate healthy donors.

Article Snippet: For cytokine immunoassay and generation of CFSE-monocytes, CD14 + PBMos were sorted from PBMCs (where specified after incubation with Liberase TL and DNase I as described in the Tissue handling and cell isolation section) with CD14 MicroBeads (Miltenyi Biotec).

Techniques: Gene Expression, In Vitro, Expressing, Isolation, Derivative Assay

SI Mfs are highly proficient at antigen uptake. (A) The percentage of pHrodo green E. coli + cells among CD14 + PBMo, PBDC (left, n = 5), and SI Mf subsets and DCs (right, n = 5) was determined by flow cytometry after indicated incubation times. (B) The effect of preincubation with human serum (opsonization) on the uptake of pHrodo green E. coli bioparticles by SI Mf subsets and DCs was determined by flow cytometry after 15 or 45 min of incubation ( n = 5). E. coli bioparticles were opsonized by incubating in 50% inactivated human serum for 30 min at 37°C. *, P < 0.05; ****, P < 0.0001 (two-way RM-ANOVA comparing effect of opsonization at one time point). (C) The percentage of DQ-OVA (green) + cells among CD14 + PBMo, PBDC (left, n = 5), and SI Mf subsets and DCs (right, n = 5) was determined by flow cytometry after indicated incubation times. (A and C) Data are presented as mean ± SEM of pHrodo + (A) or DQ-OVA + (C) cells within each population (Fig. S4 A). Comparisons between subsets are marked in color corresponding to compared subset. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 (two-way RM-ANOVA).

Journal: The Journal of Experimental Medicine

Article Title: Transcriptional and functional profiling defines human small intestinal macrophage subsets

doi: 10.1084/jem.20170057

Figure Lengend Snippet: SI Mfs are highly proficient at antigen uptake. (A) The percentage of pHrodo green E. coli + cells among CD14 + PBMo, PBDC (left, n = 5), and SI Mf subsets and DCs (right, n = 5) was determined by flow cytometry after indicated incubation times. (B) The effect of preincubation with human serum (opsonization) on the uptake of pHrodo green E. coli bioparticles by SI Mf subsets and DCs was determined by flow cytometry after 15 or 45 min of incubation ( n = 5). E. coli bioparticles were opsonized by incubating in 50% inactivated human serum for 30 min at 37°C. *, P < 0.05; ****, P < 0.0001 (two-way RM-ANOVA comparing effect of opsonization at one time point). (C) The percentage of DQ-OVA (green) + cells among CD14 + PBMo, PBDC (left, n = 5), and SI Mf subsets and DCs (right, n = 5) was determined by flow cytometry after indicated incubation times. (A and C) Data are presented as mean ± SEM of pHrodo + (A) or DQ-OVA + (C) cells within each population (Fig. S4 A). Comparisons between subsets are marked in color corresponding to compared subset. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 (two-way RM-ANOVA).

Article Snippet: For cytokine immunoassay and generation of CFSE-monocytes, CD14 + PBMos were sorted from PBMCs (where specified after incubation with Liberase TL and DNase I as described in the Tissue handling and cell isolation section) with CD14 MicroBeads (Miltenyi Biotec).

Techniques: Flow Cytometry, Incubation

SI Mfs have attenuated cytokine production. (A) Spontaneous cytokine release by sorted SI Mf subsets and DCs ( n = 8) and CD14 + PBMos and PBMo-lib ( n = 7) was measured by a Bio-Plex assay in supernatants after 21 h of culture. Bars indicate median values. # , P < 0.05; ## , P < 0.01; ### , P < 0.001; #### , P < 0.0001 (ANOVA of PBMo, PBMo-lib, and Mf1). **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 (RM-ANOVA of SI subsets). (B) Concentration of cytokines released spontaneously (dots) and after stimulation with 1 µg/ml LPS (squares) in supernatants after 21 h of culture of sorted SI Mf subsets and DCs ( n = 5). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 (two-way RM-ANOVA). (C) Concentration of cytokines released spontaneously (dots) and after stimulation with 1 µg/ml LPS (squares) in supernatants after 21 h of culture of sorted CD14 + PBMos and PBMo-lib ( n = 6 or 7). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 (two-way RM-ANOVA). (D) The percentage of TNFα + cells in SI Mf subsets was determined by intracellular staining and flow cytometric analysis after 4 h of culture in medium only or containing LPS, flagellin, R848, or poly(I:C) ( n = 3). Floating bars represent range, with a line indicating median. Dotted line represents background staining on Mf3 and Mf4 based on the fraction of cells found within the isotype control gate. *, P < 0.05; **, P < 0.01 compared with matched medium-only control (two-way RM-ANOVA).

Journal: The Journal of Experimental Medicine

Article Title: Transcriptional and functional profiling defines human small intestinal macrophage subsets

doi: 10.1084/jem.20170057

Figure Lengend Snippet: SI Mfs have attenuated cytokine production. (A) Spontaneous cytokine release by sorted SI Mf subsets and DCs ( n = 8) and CD14 + PBMos and PBMo-lib ( n = 7) was measured by a Bio-Plex assay in supernatants after 21 h of culture. Bars indicate median values. # , P < 0.05; ## , P < 0.01; ### , P < 0.001; #### , P < 0.0001 (ANOVA of PBMo, PBMo-lib, and Mf1). **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 (RM-ANOVA of SI subsets). (B) Concentration of cytokines released spontaneously (dots) and after stimulation with 1 µg/ml LPS (squares) in supernatants after 21 h of culture of sorted SI Mf subsets and DCs ( n = 5). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 (two-way RM-ANOVA). (C) Concentration of cytokines released spontaneously (dots) and after stimulation with 1 µg/ml LPS (squares) in supernatants after 21 h of culture of sorted CD14 + PBMos and PBMo-lib ( n = 6 or 7). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 (two-way RM-ANOVA). (D) The percentage of TNFα + cells in SI Mf subsets was determined by intracellular staining and flow cytometric analysis after 4 h of culture in medium only or containing LPS, flagellin, R848, or poly(I:C) ( n = 3). Floating bars represent range, with a line indicating median. Dotted line represents background staining on Mf3 and Mf4 based on the fraction of cells found within the isotype control gate. *, P < 0.05; **, P < 0.01 compared with matched medium-only control (two-way RM-ANOVA).

Article Snippet: For cytokine immunoassay and generation of CFSE-monocytes, CD14 + PBMos were sorted from PBMCs (where specified after incubation with Liberase TL and DNase I as described in the Tissue handling and cell isolation section) with CD14 MicroBeads (Miltenyi Biotec).

Techniques: Plex Assay, Concentration Assay, Staining, Control

A Representative experiment of 3 independent experiments of IAP expression levels analyzed by Western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. B Quantification of GL261-DsRed spheroids area upon vehicle ( n = 10) and SMg treatment ( n = 9) and in the presence or not of the C8B4 cells ( n = 10). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. C Concentrations of CD14 and CCL17/TARC quantified in the supernatant of C8B4 and GL261-DsRed co-cultures by ELISA assay after 72 h of vehicle or SMg treatments ( n = 6 independent experiments). Statistical analyses were performed by using Mann–Whitney test; alpha = 0.05, bilateral p-value: ** p < 0.005. D Quantification of GL261-DsRed spheroids area in the presence of the C8B4 cells expressing ML-IAP (siCTRL) or down-expressing ML-IAP (siML-IAP) ( n = 10) after 72 h of treatment vehicle ( n = 10) and SMg treatment ( n = 9). Data from three independent experiments. Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: * p < 0.05 ; ** p < 0.005 ; **** p < 0.0001. E Quantification of GL261-DsRed spheroid area in the presence of C8B4 cells treated with vehicle and SMg for 72 h. C8B4 cells were pre-treated for 24 h with ZVAD (vehicle n = 23, SMg n = 23) and TNFαi (vehicle n = 27, SMg n = 23) or without pre-treatment (vehicle n = 28, SMg n = 25). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. F Representative experiment ( n = 2) of IAP expression levels analyzed by western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. G Representative experiment ( n = 2) of expression levels of p65, phospho-p65, IκBα, phospho IκB, caspase-3, cleaved caspase-3 analyzed by Western blotting in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. H Representative experiment out of 3 experiments of expression levels of CD206 and iNOS analyzed by Western blotting after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Expression level of actin β served as loading control. I iNOS/CD206 ratio in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Quantification was performed from 3 independent experiments using ImageJ software and data presented were normalized to actin β expression. iNOS/CD206 ratio fold changes were normalized on vehicle condition. B – E , I Bar graphs represent mean ± s.e.m.

Journal: Cell Death & Disease

Article Title: SMAC mimetic drives microglia phenotype and glioblastoma immune microenvironment

doi: 10.1038/s41419-024-07056-z

Figure Lengend Snippet: A Representative experiment of 3 independent experiments of IAP expression levels analyzed by Western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. B Quantification of GL261-DsRed spheroids area upon vehicle ( n = 10) and SMg treatment ( n = 9) and in the presence or not of the C8B4 cells ( n = 10). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. C Concentrations of CD14 and CCL17/TARC quantified in the supernatant of C8B4 and GL261-DsRed co-cultures by ELISA assay after 72 h of vehicle or SMg treatments ( n = 6 independent experiments). Statistical analyses were performed by using Mann–Whitney test; alpha = 0.05, bilateral p-value: ** p < 0.005. D Quantification of GL261-DsRed spheroids area in the presence of the C8B4 cells expressing ML-IAP (siCTRL) or down-expressing ML-IAP (siML-IAP) ( n = 10) after 72 h of treatment vehicle ( n = 10) and SMg treatment ( n = 9). Data from three independent experiments. Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: * p < 0.05 ; ** p < 0.005 ; **** p < 0.0001. E Quantification of GL261-DsRed spheroid area in the presence of C8B4 cells treated with vehicle and SMg for 72 h. C8B4 cells were pre-treated for 24 h with ZVAD (vehicle n = 23, SMg n = 23) and TNFαi (vehicle n = 27, SMg n = 23) or without pre-treatment (vehicle n = 28, SMg n = 25). Statistical analyses were performed by using ANOVA test and ANOVA post-hoc Tukey test; alpha = 0.05, bilateral p -value: *** p < 0.0005. F Representative experiment ( n = 2) of IAP expression levels analyzed by western blotting after 72 h of vehicle or SMg treatment in C8B4 microglia cell line. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. G Representative experiment ( n = 2) of expression levels of p65, phospho-p65, IκBα, phospho IκB, caspase-3, cleaved caspase-3 analyzed by Western blotting in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. H Representative experiment out of 3 experiments of expression levels of CD206 and iNOS analyzed by Western blotting after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Expression level of actin β served as loading control. I iNOS/CD206 ratio in C8B4 microglia cell line after 24 h of vehicle or SMg treatment. C8B4 cells were pre-treated for 24 h with ZVAD and TNFαi. Quantification was performed from 3 independent experiments using ImageJ software and data presented were normalized to actin β expression. iNOS/CD206 ratio fold changes were normalized on vehicle condition. B – E , I Bar graphs represent mean ± s.e.m.

Article Snippet: Supernatants were collected after 72 h of treatment and processed using the proteome profiler human XL cytokine array kit (R&D systems, #ARY022B) or the ELISA kits Quantikine ELISA Mouse CCL17/TARC Immunoassay (R&D systems, MCC170) and Mouse CD14 Immunoassay (R&D systems, MC140) accordingly to the manufacturer protocol (Data represented were normalized to reference spot expression in array membrane).

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Control, Software