MiR-192-Mediated Positive Feedback Loop Controls the Robustness of Stress-Induced p53 Oscillations in Breast Cancer Cells
Fig 5
Experimental perturbation of p53-associated microRNA affects oscillatory behavior.
(a) To examine the role of microRNA 29a, 34a and 192 in the stress-induced p53 oscillations, expression of three endogenous microRNAs known to be associated with p53 activation pathway is targeted using microRNA inhibitor. Fluorescence is captured every 10 minutes for 20 hours following transfection. Single cells are identified and their intensity is recorded to create a time series data for each cell. (b) Quantitative PCR measurements of microRNA levels before and after treatment with the microRNA inhibitor. (c) Period distribution of p53 oscillation before and after treatment with various microRNA inhibitor. Magenta bars represent period of p53 oscillations observed in wild type population, and brown bars represent the same measurement from the population treated with the specified microRNA inhibitor. (d) Heatmaps showing single cell tracks of p53 fluorescence measurement after various microRNA inhibitor treatment. Each graph has 100 rows, each row representing a single cell tracking data for 20 hours. Data shown has been reorganized to reflect the results from the classification process to identify cells with oscillatory p53 expression. Number on the left shows the number of cells identified as oscillating for each group, where the magenta bar indicates the wild-type level.