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cyclin e pbi egfp  (Addgene inc)
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Addgene inc cyclin e pbi egfp
(A-D) Cells were treated with varying doses of drugs inhibiting cell cycle progression (circles) or protein production (squares). (A) The fold change in cell size (after 44 hrs in drug) vs. fold change in growth rate, relative to untreated cells, is plotted for each condition. Dotted line denotes proportional relationship expected if there is no coordination between growth and cell cycle progression. (B) The fold change in cell size (after 44 hrs in drug) vs. fold change in cell cycle length, relative to untreated cells, is plotted for each condition. (C) Mean growth rate vs. mean cell cycle length are plotted for cells in each condition, calculated from measured increases in bulk protein and number of cells over the course of a 68-hour incubation. Black lines mark iso-cell-size contours. Region between lines spans a 25% shift in cell size. (D) Cells were transfected with plasmids encoding wild-type or degradation-resistant <t>cyclin</t> E under control of a constitutive promoter (diamonds). Mean growth rate vs. mean cell cycle length are plotted for cells in each condition, overlaid on the plot shown in (C). Calculation of the Pearson correlation coefficient between log(growth rate) and log(cell cycle length), using the data shown in (D), yields r = -0.76, p = 2.1x10 −7 , by two-tailed Student’s t-test, indicating a significant inverse correlation between growth rate and cell cycle length. (E,F) Mean growth rate vs. mean cell cycle length for cells treated with drugs inhibiting cell cycle progression (E) or protein production (F), calculated for three time intervals during drug treatment: 0-14 hrs, 14-44 hrs, 44-68 hrs. Lines connect time-points in sequential order, with arrowheads pointing to latest time-point. Data is shown for 25 uM BN82002 (E, green), 1.5 uM Cdk2 Inhibitor III (E, red), 0.06 uM Cycloheximide (F, blue), 10 nM Torin (F, orange), and 7 uM Rapamycin (F, purple). For (A-F), each treatment was done in duplicate, with several thousand cells in each sample.
Cyclin E Pbi Egfp, supplied by Addgene inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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(A-D) Cells were treated with varying doses of drugs inhibiting cell cycle progression (circles) or protein production (squares). (A) The fold change in cell size (after 44 hrs in drug) vs. fold change in growth rate, relative to untreated cells, is plotted for each condition. Dotted line denotes proportional relationship expected if there is no coordination between growth and cell cycle progression. (B) The fold change in cell size (after 44 hrs in drug) vs. fold change in cell cycle length, relative to untreated cells, is plotted for each condition. (C) Mean growth rate vs. mean cell cycle length are plotted for cells in each condition, calculated from measured increases in bulk protein and number of cells over the course of a 68-hour incubation. Black lines mark iso-cell-size contours. Region between lines spans a 25% shift in cell size. (D) Cells were transfected with plasmids encoding wild-type or degradation-resistant cyclin E under control of a constitutive promoter (diamonds). Mean growth rate vs. mean cell cycle length are plotted for cells in each condition, overlaid on the plot shown in (C). Calculation of the Pearson correlation coefficient between log(growth rate) and log(cell cycle length), using the data shown in (D), yields r = -0.76, p = 2.1x10 −7 , by two-tailed Student’s t-test, indicating a significant inverse correlation between growth rate and cell cycle length. (E,F) Mean growth rate vs. mean cell cycle length for cells treated with drugs inhibiting cell cycle progression (E) or protein production (F), calculated for three time intervals during drug treatment: 0-14 hrs, 14-44 hrs, 44-68 hrs. Lines connect time-points in sequential order, with arrowheads pointing to latest time-point. Data is shown for 25 uM BN82002 (E, green), 1.5 uM Cdk2 Inhibitor III (E, red), 0.06 uM Cycloheximide (F, blue), 10 nM Torin (F, orange), and 7 uM Rapamycin (F, purple). For (A-F), each treatment was done in duplicate, with several thousand cells in each sample.

Journal: bioRxiv

Article Title: Cell size sensing in animal cells coordinates anabolic growth rates with cell cycle progression to maintain uniformity of cell size

doi: 10.1101/123851

Figure Lengend Snippet: (A-D) Cells were treated with varying doses of drugs inhibiting cell cycle progression (circles) or protein production (squares). (A) The fold change in cell size (after 44 hrs in drug) vs. fold change in growth rate, relative to untreated cells, is plotted for each condition. Dotted line denotes proportional relationship expected if there is no coordination between growth and cell cycle progression. (B) The fold change in cell size (after 44 hrs in drug) vs. fold change in cell cycle length, relative to untreated cells, is plotted for each condition. (C) Mean growth rate vs. mean cell cycle length are plotted for cells in each condition, calculated from measured increases in bulk protein and number of cells over the course of a 68-hour incubation. Black lines mark iso-cell-size contours. Region between lines spans a 25% shift in cell size. (D) Cells were transfected with plasmids encoding wild-type or degradation-resistant cyclin E under control of a constitutive promoter (diamonds). Mean growth rate vs. mean cell cycle length are plotted for cells in each condition, overlaid on the plot shown in (C). Calculation of the Pearson correlation coefficient between log(growth rate) and log(cell cycle length), using the data shown in (D), yields r = -0.76, p = 2.1x10 −7 , by two-tailed Student’s t-test, indicating a significant inverse correlation between growth rate and cell cycle length. (E,F) Mean growth rate vs. mean cell cycle length for cells treated with drugs inhibiting cell cycle progression (E) or protein production (F), calculated for three time intervals during drug treatment: 0-14 hrs, 14-44 hrs, 44-68 hrs. Lines connect time-points in sequential order, with arrowheads pointing to latest time-point. Data is shown for 25 uM BN82002 (E, green), 1.5 uM Cdk2 Inhibitor III (E, red), 0.06 uM Cycloheximide (F, blue), 10 nM Torin (F, orange), and 7 uM Rapamycin (F, purple). For (A-F), each treatment was done in duplicate, with several thousand cells in each sample.

Article Snippet: Cyclin E-pBI-EGFP was a gift from Bert Vogelstein (Addgene plasmid # 16654)( Rajagopalan, H. et al.

Techniques: Incubation, Transfection, Control, Two Tailed Test