Breast Cancer Cell Migration Assay

MCF7, SK-BR-3 and MDA-MB-231 breast cancer cell lines were starved over night in serum-free DMEM medium. Breast cancer cells (1×10⁵) in DMEM with 2% FBS were added to the top of each Boyden migration chamber (8 μm, 12-well plate format; BD Biosciences, Palo Alto, CA) with or without CSF-1R-blocking antibody (10μg/ml) followed by treatment with 200 ng/ml recombinant IL-34 or CSF-1 protein (R&D Systems, McKinley Place, MN, USA). After 48 h, medium was removed and membranes were processed as described [59 (link)].

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Zins K., Heller G., Mayerhofer M., Schreiber M, & Abraham D. (2018). Differential prognostic impact of interleukin-34 mRNA expression and infiltrating immune cell composition in intrinsic breast cancer subtypes. Oncotarget, 9(33), 23126-23148.

Publication 2018

Boyden migration chamber recombinant IL-34

Blocking antibody Breast cancer Cells Csf 1 Il 34 Mcf7 Mda mb 231 cell lines Membranes Protein Serum

Corresponding Organization : Comprehensive Cancer Center Vienna

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Variable analysis

independent variables

CSF-1R-blocking antibody (10μg/ml)
200 ng/ml recombinant IL-34
200 ng/ml recombinant CSF-1 protein

dependent variables

Cell migration through the Boyden migration chamber

control variables

MCF7, SK-BR-3 and MDA-MB-231 breast cancer cell lines
Serum-free DMEM medium for starvation
DMEM with 2% FBS for cell culture
Boyden migration chamber (8 μm, 12-well plate format)
Incubation time of 48 h

controls

Positive control: Cells treated with CSF-1 protein
Negative control: Cells without any treatment

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