supp_figure4d: CCND1 protein expression in PC3 stable lines.

In GranderLab/miR34a_asRNA_project: The miR34AasRNAproject package facilitates the transparency and reproducibility of the miR34a asRNA project and associated publication.

Description

All cell lines were cultured at 5 (Hyclone) and 2 mM L-glutamine. All growth mediums were supplemented with 10 penicillin. We seeded three independent experiments of 150,000 cells/well in a six well plate (PC3 mock vs. PC3 miR34a AS F4) and grew them for 24 hours. Before harvesting cells were controlled for their confluency. Cells with a confluence of 60-75 down at -80<c2><b0>C before lysed for Western Blot analysis. Cells were lysed in RIPA Buffer and run on a 4-12 Proteins were transferred onto a nitrocellulose membrane using iBlot Turbo Blotting Device. Proteins were transferred for 7 minutes. Membranes were blocked for 1 hour at room temperature in 5 with a cyclin D1 antibody (92G2 Rabbit mAb, Cell Signalling) overnight at 4<c2><b0>C. The membrane was incubated using a goat anti-rabbit antibody conjugated to HRP for 1 hour at room temperature. Membranes were washed 3-times for 5min each in TBS-T. Membranes were developed using chemiluminescence. After the picture was taken, the membrane was stripped using 0.4M NaOH solution for 20min. Blocking step was repeated and primary antibody against GAPDH was incubated for overnight at 4<c2><b0>C. Consequently, the membrane was incubated using a goat anti-rabbit antibody conjugated to HRP for 1 hour at room temperature. Membranes were washed 3-times for 5min in TBS-T. Membranes were developed using chemiluminescence.

Format

A tibble with 12 rows and 6 variables:

'Biological Replicate'

Numeric; Indicates the biological replicate of the sample.

condition

Factor; Indicates the genetic modifications in the cell line.

protein

Character; indicates the protein that the values correspond to.

pixel

Numeric; https://imagej.nih.gov/ij/.

Intensity

Numeric; https://imagej.nih.gov/ij/.

background

Numeric; https://imagej.nih.gov/ij/.

...

Examples

getData('Supplementary Figure 4d')

GranderLab/miR34a_asRNA_project documentation built on May 26, 2019, 7:26 a.m.