Supplementary materials

Real-time quantification of single RNA translation dynamics in living cells

Tatsuya Morisaki, Kenneth Lyon, Keith F. DeLuca, Jennifer G. DeLuca, Brian P. English, Zhengjian Zhang, Luke D. Lavis, Jonathan B. Grimm, Sarada Viswanathan, Loren L. Looger, Timothee Lionnet, Timothy J. Stasevich

Materials/Methods, Supplementary Text, Tables, Figures, and/or References

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  • Materials and Methods
  • Supplementary Text
  • Figs. S1 to S12
  • Full Reference List
  • Captions for Movies S1 to S8

Images, Video, and Other Other Media

Movie S1
Max projection of a 13 z-stack movie showing sample SM-KDM5B polysomes imaged at one frame per second for 120 (of 300) seconds total (Ch1 red, mRNA; Ch2 green, protein).
Movie S2
Max projection of a 13 z-stack movie showing sample SM-KDM5B polysomes imaged at one frame per 1 minute for 2 hours total (Ch1 red, mRNA; Ch2 green, protein).
Movie S3
Max projection of a 13 z-stack movie of a puromycin experiment showing a loss of protein signal at SM-KDM5B polysomes compared to control cells. Cell imaged at one frame per 6.5 seconds for a total of 650 seconds (Ch1 red, mRNA; Ch2 green, protein).
Movie S4
Max projection of a 13 z-stack movie showing sample SM-beta-actin polysomes imaged at one frame per second for 120 (of 300) seconds total (Ch1 red, mRNA; Ch2 green, protein).
Movie S5
Max projection of a 13 z-stack movie showing sample SM-H2B polysomes imaged at one frame per second for 120 (of 300) seconds total (Ch1 red, mRNA; Ch2 green, protein).
Movie S6
Animated GIF of a sample stochastic Monte Carlo simulation movie of translation of SM-KDM5B transcripts. The green box shows spaghetti monster tag coding region, and the black box shows KDM5B coding region. The green circle above shows the total fluorescence intensity that would be measured at the translation site. The fluorescence time series is shown below. The x-axis is in minutes and the y-axis is in arbitrary fluorescence intensity units.
Movie S7
Animated GIF of a sample stochastic Monte Carlo simulation movie of 100 μg/ml cycloheximide stalled translation of SM-KDM5B transcripts. The green box shows spaghetti monster tag coding region, and the black box shows KDM5B coding region. The green circle above shows the total fluorescence intensity that would be measured at the translation site. The fluorescence time series is shown below. The x-axis is in minutes and the y-axis is in arbitrary fluorescence intensity units.
Movie S8
Max projection of a 13 z-stack movie showing a sample cell with two types of polysomes, FLAG-KDM5B and HA-KDM5B, labeled by Fab against the FLAG and HA tags, respectively. Cell imaged at one frame every 2 seconds for 200 seconds total (Ch1 red, mRNA; Ch2 green, FLAG; Ch3 blue, HA).