Imaging endogenous RNA polymerase II phosphorylation at a single copy gene in living cells.
Автор: Timothy Stasevich Lab
Загружено: 2021-05-10
Просмотров: 482
Описание: Following the transcription cycle of endogenous RNA polymerase II at a single copy gene in living cells. Maximum projection of a 13 z-stack three-color movie showing an exemplary Hela-128 cell. Transcription at a single-copy HIV-1 reporter gene (circled) is highlighted. At the transcription site, mRNA can be see in blue, and endogenous RNA polymerase II (RNAP2) can be see in red (pan-RNAP2) and green (RNAP2 phosphorylated at Serine 5). Raw and bandpass filtered crops for each signal and a merge (white) over time are shown in the insets. Images were acquired every 1 min for a total of 200 min, shown here as a moving average (over three time points). At select time points, various signals can be seen in the absence of the others, confirming no fluorescence bleed through, and demonstrating signals are not perfectly synchronous. For example, t=30-40 min shows a strong burst of all three signals, while t=162-165 min shows CTD-RNAP2 without Ser5ph-RNAP2, t=51-54 min shows Ser5ph-RNAP2 without CTD-RNAP2, and t=149-154 min shows mRNA without the other RNAP2 signals. Scale bar, 10 µm.
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