non-linear regression analysis (sigmoidal, 4pl)  (GraphPad Software Inc)

Journal: Cell chemical biology

Article Title: The Natural Product Butylcycloheptyl Prodiginine Binds Pre-miR-21, Inhibits Dicer-Mediated Processing of Pre-miR-21, and Blocks Cellular Proliferation

doi: 10.1016/j.chembiol.2019.04.011

Figure Lengend Snippet: Differential scanning fluorimetry and cytotoxicity results for pre-miR-21 stability modulating compounds. PubChem CID # is the compound’s identifier ( https://pubchem.ncbi.nlm.nih.gov/ ). The Δ T m MAX was calculated by taking the melting temperature shift difference of pre-miR-21 treated with the compound vs untreated. a The apparent Kd ( K d app ) was measured by direct binding assay using intrinsic fluorescence max 545/580 nm. b GI 50 was determined using the XTT cell viability assay. Calculations were performed using the non-linear regression fit (Sigmoidal, 4PL) function in GraphPad Prism. cNormal colon cells used were ATCC CRL-1790. d 1,5-bis[3-(2-hydroxyethylamino)propylamino]anthracene-9,10-dione hydrochloride. All experiments were repeated in triplicate.

Article Snippet: T m was obtained using the non-linear regression fit (Sigmoidal, 4PL) function in GraphPad Prism where y = fluorescence, x = temperature. ( C ) a concentration of 0, 12.5, 25, 50, and 100 nM of 1 was incubated with buffer (black), 50 ng/μL purified total RNA from HCT-116 cells (red), 25 ng/μL purified genomic DNA from HCT-116 cells (green), or 50 ng/μL of plasmid DNA (blue).

Techniques: Binding Assay, Fluorescence, Viability Assay