Immunofluorescence Staining of Tissue Samples

Immunofluorescence staining was performed on 6-µm-thick frozen or paraffin-embedded tissue sections, as previously described (31 (link)). The following antibodies were used: Mouse monoclonal antibodies against TGF-β2 (#ab36495), vimentin (#ab8059) and 5-methylcytosine (#ab10805) (all from Abcam), and rabbit monoclonal antibodies against E-cadherin (#3195s; Cell Signaling Technology, Inc.) and Ki-67 (#ab92742; Abcam). Primary antibodies were diluted 1:50 and secondary antibodies 1:400; the incubations were all performed at room temperature. Signals were detected with a Leica DMRB fluorescence microscope (Leica Microsystems Ltd., Milton Keynes, UK). Images of representative fields were captured using a SPOTTM FLEX 15.2 64-Mp shifting pixel digital color camera and analyzed with SPOT Basic/Advanced 4.6 software (both from Diagnostic Instruments, Sterling Heights, Michigan, USA).

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Abukiwan A., Nwaeburu C.C., Bauer N., Zhao Z., Liu L., Gladkich J., Gross W., Benner A., Strobel O., Fellenberg J, & Herr I. (2018). Dexamethasone-induced inhibition of miR-132 via methylation promotes TGF-β-driven progression of pancreatic cancer. International Journal of Oncology, 54(1), 53-64.

Publication 2018

ab36495 ab10805 E-cadherin ab92742 Leica DMRB fluorescence microscope

5 methylcytosine Antibodies Cadherin Diagnostic Flex Fluorescence microscope Frozen Immunofluorescence Monoclonal antibodies Mouse Paraffin Rabbit Tgf β2 Tissue Vimentin

Corresponding Organization :

Other organizations : Heidelberg University, German Cancer Research Center

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Variable analysis

independent variables

Tissue section thickness (6-µm)
Tissue source (frozen or paraffin-embedded)

dependent variables

Immunofluorescence staining patterns for TGF-β2, vimentin, 5-methylcytosine, E-cadherin, and Ki-67

control variables

Antibody dilutions (primary antibodies: 1:50, secondary antibodies: 1:400)
Incubation conditions (all performed at room temperature)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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