prism encoded tests  (GraphPad Software Inc)

Journal: Scientific Reports

Article Title: Aspergillus Carneus metabolite Averufanin induced cell cycle arrest and apoptotic cell death on cancer cell lines via inducing DNA damage

doi: 10.1038/s41598-023-30775-w

Figure Lengend Snippet: γ-H2Ax staining showed accumulation of DNA damage upon Averufanin treatment. Human breast cancer cells (T47D) were treated with ( A ) DMSO, ( B ) IC 50 dose Averufanin, or ( C ) IC 75 dose Averufanin for 48 h and stained with anti-γ-H2Ax and DAPI. Blue represents DAPI and red represents γ-H2Ax which increases as a DNA damage response (DDR). Images were obtained from the confocal microscopy through LasX and dye intensities were evaluated with ImageJ. H2A.X intensities were normalized to DAPI, graphics and statistical analysis were conducted with GraphPad PRISM. Two-tailed, unpaired t-test was applied between DMSO Control and treatment groups (*p < 0.05).

Article Snippet: Statistical analysis was performed with GraphPad PRISM encoded tests.

Techniques: Staining, Confocal Microscopy, Two Tailed Test

Journal: Scientific Reports

Article Title: Aspergillus Carneus metabolite Averufanin induced cell cycle arrest and apoptotic cell death on cancer cell lines via inducing DNA damage

doi: 10.1038/s41598-023-30775-w

Figure Lengend Snippet: Cellular pathway components targeted by Averufanin treatment. Human breast cancer cells (T47D) were treated with the IC 50 and IC 75 concentrations of Averufanin or DMSO control for 48 h. ( A ) Western blot analysis showed that p53 protein was activated by phosphorylation upon treatment with Averufanin. Furthermore, ( B ) PARP protein was shown to be cleaved indicating induction of apoptosis. ( C ) pGSK3β and ( D ) Cyclin D levels were decreased in correlation with the cell cycle analyses. ( E ) Cyclin A2 and ( F ) CDK2 levels were shown to be increased in correlation with DNA damage response. Original blots are presented in Supplementary Figures. Quantification of the results were obtained using ImageJ software. Graphics and statistical analysis were performed with GraphPad PRISM, DMSO vs Treatment groups and p values are in the brackets. ( G ) Schematic representation of the molecular mechanism of action signaling of the compounds. Blocked pathways are crossed. Multi-step downstream effects are represented with dashed arrows. γ-H2Ax staining results indicated DNA damage caused by the compound, followed by dephosphorylation of pGSK3β and phosphorylation of p53. DNA damage induced activation of tumor suppressor p53 activates Caspase 3/7 pathway followed by PARP cleavage, along with CDK2-Cyclin A2 cell cycle proteins for DNA repair. At the same time Cyclin D1 is inhibited to stop the cell cycle until DNA damage can be repaired. (***p < 0.001 **p < 0.01, *p < 0.05 ns p < 0.5).

Article Snippet: Statistical analysis was performed with GraphPad PRISM encoded tests.

Techniques: Western Blot, Software, Staining, De-Phosphorylation Assay, Activation Assay