Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: New insights in gene expression alteration as effect of doxorubicin drug resistance in triple negative breast cancer cells

doi: 10.1186/s13046-020-01736-2

Figure Lengend Snippet: Evaluation of the CXCL1, IL-6 and TNF-α released in cell culture medium by ELISA, for parenteral cells (P0), P12 and P24 for Hs578T and MBA-MB-231 cell lines

Article Snippet: For TNF-α was used Human TNF-α DuoSet ELISA (R&D System, cat no. D210 ), and IL-6 DuoSet ELISA (R&D System, cat no. DY206) for IL-6 quantification along with DuoSet Ancillary Reagent Kit 2 (R&D Systems, cat no. DY008).

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Cell death & disease

Article Title: Synovial mesenchymal progenitor derived aggrecan regulates cartilage homeostasis and endogenous repair capacity.

doi: 10.1038/s41419-022-04919-1

Figure Lengend Snippet: Fig. 3 Aggrecan secretion. Aggrecan secretion from normal and OA MPCs was analyzed by ELISA and it was found that OA MPCs secreted significantly less aggrecan than normal MPCs at all time points examined (A). To test if this phenotype was in part due to the inability of aggrecan to leave the ER/Golgi complex, normal (B) or OA (C) MPCs were treated with Brefeldin A (BFA). BFA treatment of normal MPCs significantly decreased aggrecan expression (B), while BFA treatment had no effect on aggrecan secretion in OA MPCs (C). To control for a difference in gene expression, mRNA in normal and OA MPCs was quantified, and no difference was observed (D). Flow cytometry (E–G) and immunofluorescence (H, I) was used to examine if BFA treatment of normal MPCs resulted in the same aggrecan staining phenotype observed in OA cells. BFA treatment resulted in a shift to high-positive gate (E, F) that was reversible with removal of BFA (G) and the intense aggrecan staining adjacent to the nucleus was also observed with BFA treatment (I). Scale bars equal 10 µm. *p < 0.05.

Article Snippet: HA was also measured in cell culture supernatant using the Hyaluronan DuoSet ELISA (R&D Systems, Cat# DY361405) according to the manufacturer’s protocol.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Control, Gene Expression, Flow Cytometry, Staining

Journal: Clinical & Translational Immunology

Article Title: Immune activation and response dynamics of human iPSC ‐derived macrophages in tuberculosis infection models

doi: 10.1002/cti2.70071

Figure Lengend Snippet: iMacs demonstrate enhanced phagocytosis and cytokine production upon BCG infection. Microscopic analysis of immunofluorescent images stained for (a) vATPase and (b) LAMP‐1 immunolabelling after infection with BCG (MOI 10:1) for various time points, using a Leica TCS SP5 confocal microscope (100× oil objective, scale bar = 10 μm). Right panels (a) + (b) show Pearson correlation coefficient demonstrating correlation between (a) BCG (green) and v‐APTase (red) accumulation and (b) BCG (green) and LAMP‐1 (red) accumulation at different time points post‐infection. The data are representative of n = 3 independent experiments and individual cells were quantified according to the program. (c) IL‐6, (d) TNF and (e) IL‐1β secretion by iMacs and MDMs 6 h and 24 h post BCG infection with an MOI of 10:1, compared to non‐infected cells as controls measured by ELISA. Data (c) – (e) are represented as the mean ± SD of n = 4–5 biological replicates measured in duplicates. Statistical significance was determined using Mann–Whitney U test (* indicates P < 0.0332, ** indicates P < 0.0021); n.i., non‐infected control cells.

Article Snippet: ELISAs were performed using the Human IL‐6 DuoSet ELISA (R&D Systems, Cat. No. DY206) and the TNF‐alpha DuoSet ELISA (R&D Systems, Cat. No. DY210), following the manufacturer's instructions.

Techniques: Infection, Staining, Microscopy, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Control

Journal: Clinical & Translational Immunology

Article Title: Immune activation and response dynamics of human iPSC ‐derived macrophages in tuberculosis infection models

doi: 10.1002/cti2.70071

Figure Lengend Snippet: iMacs exhibit a beneficial immune response against HKMT . (a) Representative phase‐contrast microscopy images of Selene iMacs (left), NIH iMacs (middle) and MDMs (right) non‐stimulated or post‐HKMT stimulation using a Leica TCS SP5 confocal microscope (scale bars = 100 μm top and 200 μm bottom) ( n = 2). (b) Flow cytometric quantification of the percentage of phagocytic macrophages at different time points post‐HKMT stimulation ( n = 8–13). (c) ELISA quantification of IL‐6 and (d) TNF secretion 24 h post HKMT stimulation compared with non‐stimulated cells as controls ( n = 4–20). (e) Cytokine bead assay analysis of additional pro‐ and anti‐inflammatory cytokines 24 h post HKMT stimulation ( n = 3–6). (f) Flow cytometric quantification of ROS production upon HKMT stimulation. Statistical significance is indicated using a two‐tailored t ‐test ( n = 9–13). (g) RT‐qPCR analysis of Bax expression, (h) Cathepsin B expression and (i) LC3B expression in iMacs and MDMs post‐HKMT stimulation, presented as fold change relative to non‐infected cells as control ( n = 3–6). All data are presented as mean ± SD. All statistical significance was determined using a Mann–Whitney U test (* indicates P < 0.0332, ** indicates P < 0.0021, *** indicates P < 0.0002 and **** indicates P < 0.0001). n.s., non‐stimulated control cells.

Article Snippet: ELISAs were performed using the Human IL‐6 DuoSet ELISA (R&D Systems, Cat. No. DY206) and the TNF‐alpha DuoSet ELISA (R&D Systems, Cat. No. DY210), following the manufacturer's instructions.

Techniques: Microscopy, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Expressing, Infection, Control, MANN-WHITNEY