recombinant human mif rmif  (MedChemExpress)

Journal: bioRxiv

Article Title: MIF Regulates M1 Macrophage Polarization via CD74/CXCR2/JNK Pathway and Mediates Aortic Dissection in Mice

doi: 10.1101/2023.10.26.564292

Figure Lengend Snippet: RAW264.7 cells pretreated with ISO-1 (10-50μmol/L) and stimulated with LPS/IFN-γ for 24h. A , Western blot images and quantification of MIF, iNOS and IL-18 in RAW264.7 cells. B , Representative flow cytometry charts of RAW264.7 cells in each group and quantification of M1 macrophages (CD86+CD206-). C , Cytotoxic effects of rMIF (0-1000ng/ml) on RAW264.7 cells assessed by CCK8 assay. D , Administration of 0-500 ng/ml rMIF stimulated RAW264.7 cells for 15min-24h. Western blot was used to analyze the expression of iNOS and IL-18 proteins, and the protein expression of each group was quantified using the 0ng/ml group as a control.

Article Snippet: Macrophages were stimulated with rMIF (MCE, USA) at concentrations of 10, 50, 100 and 500 ng/ml for 15 min, 30 min, 1h, 6h, 12h and 24h, and cells were collected for relevant experiments.

Techniques: Western Blot, Flow Cytometry, CCK-8 Assay, Expressing

Journal: bioRxiv

Article Title: MIF Regulates M1 Macrophage Polarization via CD74/CXCR2/JNK Pathway and Mediates Aortic Dissection in Mice

doi: 10.1101/2023.10.26.564292

Figure Lengend Snippet: A , Western blot images and quantification of iNOS and IL-18 in RAW264.7 cells treated by 50ng rMIF for 15min-24h. B , RT-qPCR of iNOS, TNF-α, IL-6 and Arg-1 in RAW264.7 cells treated by 0-500 ng/ml rMIF. C , Representative photographs and quantification of immunofluorescence staining of iNOS (green) and DAPI (blue) in RAW264.7 cells treated with MIF or LPS/IFN-γ. D , Representative flow cytometry charts of RAW264.7 cells treated with MIF or LPS/IFN-γ and quantification of M1 macrophages (CD86+CD206-).

Article Snippet: Macrophages were stimulated with rMIF (MCE, USA) at concentrations of 10, 50, 100 and 500 ng/ml for 15 min, 30 min, 1h, 6h, 12h and 24h, and cells were collected for relevant experiments.

Techniques: Western Blot, Quantitative RT-PCR, Immunofluorescence, Staining, Flow Cytometry

Journal: Science advances

Article Title: A MYCN-independent mechanism mediating secretome reprogramming and metastasis in MYCN -amplified neuroblastoma.

doi: 10.1126/sciadv.adg6693

Figure Lengend Snippet: Fig. 6. MIF secretion enhanced by MYO1B promotes NB cell invasion and metastasis. (A) Evaluation of MIF concentrations by ELISA in unprocessed conditioned medium (CM) from the indicated siRNA-transfected Kelly cells (n = 4). (B and C) Impact of filtration by filters with 100 kDa or 20-nm pore sizes (B) or RIPA + sonication treatment (C) on MIF concentrations in CM samples from Kelly cells (n = 4). (D) Impact of MIF or MYO1B depletion on MIF concentrations in RIPA + sonication–treated CM samples from Kelly cells (n = 3). (E) Colocalization of MIF with MYO1B in Kelly cells was evaluated by IF confocal microscopy. (F and G) Impact of MIF or MYO1B depletion ± recombinant MIF (rMIF) treatment on cell invasion through Matrigel was evaluated by Incucyte (n = 5 to 8). (H and I) Impact of MIF depletion on the extravasation (H) and metastatic capacity (I) of luciferase-expressing Kelly cells was evaluated using the chick embryo CAM metastasis model. The metastasis burden was measured by BLI. Differences between groups were determined by two-tailed unpaired Student’s t test. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: For rMIF treatment (catalog no. 289-MF, R&D Systems), rMIF was added to the Matrigel at 0, 20, or 100 ng/ml.

Techniques: Enzyme-linked Immunosorbent Assay, Transfection, Filtration, Sonication, Confocal Microscopy, Recombinant, Luciferase, Expressing, Two Tailed Test