Protocol detail

Immunofluorescence Analysis of Ki67 Expression

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IF analysis was performed as described [31 (link)]. Cells grown on cover-glasses were fixed in 4% paraformaldehyde, permeabilized with 0.3% Triton X-100, and blocked with 5% bovine serum. Then the cells were incubated with antibodies against Ki67 (1:500, Abcam, USA) overnight at 4°C, followed by Alexa Fluor 594-conjugated secondary antibodies (1:500, Abcam, USA). Nuclei were dyed by DAPI and images were observed under a fluorescence microscope (Leica, Germany).

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Hu X., Wang P., Qu C., Zhang H, & Li L. (2021). Circular RNA Circ_0000677 promotes cell proliferation by regulating microRNA-106b-5p/CCND1 in non-small cell lung cancer. Bioengineered, 12(1), 6229-6239.

Publication 2021

Alexa Fluor 594-conjugated secondary antibodies fluorescence microscope

Alexa 594 Antibodies Bovine Cells Dapi Fluorescence microscope Glasses Nuclei Paraformaldehyde Serum Triton x 100

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Variable analysis

independent variables

Antibodies against Ki67 (1:500, Abcam, USA)

dependent variables

Ki67 expression

control variables

Cells grown on cover-glasses
Cells fixed in 4% paraformaldehyde
Cells permeabilized with 0.3% Triton X-100
Cells blocked with 5% bovine serum
Nuclei dyed by DAPI

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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