The lower scores obtained for combinations between taxanes and Pt drugs might be linked to the reverse mode of the screening assay, as the mode of action of mitotic inhibitors may vary for attached cells and for cell suspensions and in some cases even from the concentration used. summary of the screening experimental protocol. I) Addition of PCL Sodium lauryl sulfate compounds (10 M, 1 well/compound, 320 compounds/screening plate), positive control (10 M Doxorubicin.HCl, last two columns of every screening plate) in green and negative control (0.1% DMSO, first two columns of every screening plate) in light blue; II) Addition of cell suspension, followed by addition of Pt drugs in medium (PCL+Pt plates) or PBS/water in medium (PCL only plates) in every well of the respective plate; III) Cell viability determination by means of the Presto Blue assay after the respective drug exposure occasions; IV) Data processing, management and statistical validation; identification of HCs. All conditions were assayed in duplicate.(TIFF) pone.0211268.s003.tiff (3.1M) GUID:?A055021B-E52F-43E1-A089-07C0B32ACE10 S4 Fig: HCs (above the additive line) in A) PANC-1 and B) A549 cells identified during the primary screening. HTS Scores from the combination (i.e., PCL+Pt drugs) plates are plotted Sodium lauryl sulfate Sodium lauryl sulfate vs. the scores obtained from the PCL alone plates. Scores are given as mean from 2 replicates (2 wells/drug, respectively drug combination).(TIFF) pone.0211268.s004.tiff (690K) GUID:?68A3BDEF-571E-4DF6-9110-E34064C10785 S5 Fig: HTS scores of daunorubicin. HCl alone and in combination with cisplatin or carboplatin obtained at the confirmation screening low concentration setting in PANC-1 cells. Data is usually presented as mean SD from 2 replicates (**p < 0.01, determined by unpaired t test with Welchs correction using GraphPad Prism 7). Chemical structures (right).(TIFF) pone.0211268.s005.tiff (614K) GUID:?77034842-9F50-4049-B346-D54F9BA970D3 S6 Sodium lauryl sulfate Fig: HTS scores of chosen antimetabolites (10 Rabbit polyclonal to EGR1 M) alone and in combination with cisplatin (7 M) obtained at the confirmation screening in A549 cells. Data is usually presented as mean SD from 2 replicates (**p < 0.01 and *p < 0.05, determined by multiple (unpaired) t-tests with GraphPad Prism 7). Chemical structures (right).(TIFF) pone.0211268.s006.tiff (522K) GUID:?92BA59EA-7293-4679-9473-E79D83E294C9 S7 Fig: Chemical formula of the antihypertensive drug spironolactone. (TIF) pone.0211268.s007.tif (117K) GUID:?E07C86F4-1648-42D9-9FD4-1DC26A10907C S8 Fig: Synergistic combinations of carboplatin in PANC-1 cells. Left: Fa-CI plot of Chou-Talalay for carboplatin + topotecan (1 : 0.08; in blue), carboplatin + aminacrine (1 : 0.03; in red) and carboplatin + hycanthone (1 : 0.10; in green) Sodium lauryl sulfate combinations (72 h of exposure). Error bars represent 95% confidence intervals of the CI variability at the presented effect levels, as determined by S.D.A. Right: Chemical formulas.(TIFF) pone.0211268.s008.tiff (447K) GUID:?5951B3BD-B948-4EE4-9C94-C6158A409FCE S9 Fig: Synergistic combination of cisplatin and vorinostat (1 : 0.75) in PANC-1 cells. Left: classical isobologram at 0.5, 0.7 and 0.9 effect level (IC50, IC75 and IC90 concentrations, respectively). Markers for the actual combination points are pattern packed. Right: Chemical formulas.(TIFF) pone.0211268.s009.tiff (322K) GUID:?68DBDA47-BCB1-4F5F-8AB9-BBF8F7C794EA S10 Fig: Synergistic combinations between cisplatin and antimetabolites in A549 cells. Classical isobolograms at 0.5, 0.7 and 0.9 effect level (IC50, IC75 and IC90 concentrations, respectively) for the combinations of cisplatin with azacytidine-5, 1 : 3.72 (left) and cisplatin with gemcitabine, 1 : 0.01 (right). Markers for the actual combination points are pattern packed.(TIFF) pone.0211268.s010.tiff (395K) GUID:?4F6C2F5B-FFE1-436A-94F6-D739FABD1C31 S11 Fig: Concentration effect curves of oxaliplatin, alone and in combination with corticosteroids: Prednicarbate (at fixed concentration of 0.1 M) and flumethasone (at fixed concentration of 1 1.6 M) in A549 cells after 72 h of exposure. Curves fitting and graphs are prepared with GraphPad Prism 7.(TIFF) pone.0211268.s011.tiff (229K) GUID:?46A71C15-4533-47D3-816D-D06830C43BB9 S12 Fig: Combination effects evaluated by the fractional product method of Webb. A-D): additive to antagonistic interactions between Pt drugs and corticosteroids or paclitaxel in A549 cells; E-F): synergistic interactions between cisplatin and vorinostat, and.