squared pearson correlation coefficient (r 2)  (GraphPad Software Inc)

Journal: NPJ Breast Cancer

Article Title: Mutant P53 induces MELK expression by release of wild-type P53-dependent suppression of FOXM1

doi: 10.1038/s41523-019-0143-5

Figure Lengend Snippet: Western blotting analysis of MELK and p53 protein levels in p53-null cells (MDA MB 436 ( a ) and SKOV3 ( c )) after inducing wild-type p53 for 48 h. Wild-type p53 was induced using a doxycycline-inducible system, in p53-null cells. p21 expression was used to demonstrate the functional activity of wild-type p53 and GAPDH was used as a loading control. Numbers 1, 2, and 3 indicates triplicates of protein samples in a , c . MELK protein levels from a and c were normalized to GAPDH levels and were plotted for MDA MB 436 ( b ) and SKOV3 ( d ) cells. e Western blotting analysis of MELK, p21, and p53 protein levels in MCF7 and Cal51 cells after inducing wild-type p53 with doxorubicin (“Doxo”) at increasing concentrations (0, 10, and 25 nM) for 48 h. f Analysis of breast cancer dataset to determine the correlation between MELK mRNA levels and wild-type p53-repressed genes in the Curtis breast cancer dataset. The scale for the expression of MELK ( X -axis) and p53-repressed genes ( Y -axis) is log2-median-centered ratio. The Pearson’s correlation p -values and r 2 -values were calculated using GraphPad prism. *Statistical significance of p -value < 0.05. Error bars represent ± SD.

Article Snippet: Correlation curves and Pearson’s p -values and r 2 -values were generated using GraphPad Prism.

Techniques: Western Blot, Expressing, Functional Assay, Activity Assay, Control

Journal: NPJ Breast Cancer

Article Title: Mutant P53 induces MELK expression by release of wild-type P53-dependent suppression of FOXM1

doi: 10.1038/s41523-019-0143-5

Figure Lengend Snippet: a Western blotting analysis of MELK and FOXM1 protein levels in TNBC cells (MDA 468 and HCC70) after knocking down FOXM1. b Western blotting analysis of FOXM1 protein levels after wild-type p53 knockdown in MCF7 and ZR-75 cells. c Western blotting analysis of MELK and p53 protein levels in p53-null cells, SKOV3 (left) and MDA MB 436 (right) after inducing wild-type p53 for 48 h. Wild-type p53 was induced, using a doxycycline-inducible system, in p53-null cells. d Correlation analysis of MELK mRNA levels with FOXM1 and AURKB (FOXM1 downstream gene) in TCGA breast cancer dataset. The scale for the expression of MELK, FOXM1, and AURKB is log2-median-centered ratio. The Pearson’s correlation p -values and r 2 -values were calculated using GraphPad prism. e Western blotting analysis of MELK, FOXM1, and E2F1 protein levels in p53-mutant cells (MDA 436 and MDA 468) and wild-type p53 cells (MCF7) after E2F1 knocking down. f Western blotting analysis of E2F1 in MDA 436 cells after inducing wild-type p53 expression using doxycycline (Dox)-inducible system. g E2F1 ChIP assay to determine the relative recruitment of E2F1 to FOXM1 promoter in p53 WT cells (MCF7 and Cal51) and p53 mutant cells (MDA MB 436 and MDA MB 468). Triplicates of ChIP samples were subjected to qPCR assay using primers against FOXM1 promoter (see Supplementary Table for primers). The qPCR data were normalized to % input and then relative recruitment of E2F1 was calculated as a fold change over IgG. *Statistical significance of p -value < 0.05. Error bars represent ± SD.

Article Snippet: Correlation curves and Pearson’s p -values and r 2 -values were generated using GraphPad Prism.

Techniques: Western Blot, Knockdown, Expressing, Mutagenesis