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Maximum-intensity 2D projections of 3D image stacks calculated for each 1-min time point. Time given in minutes. Scale bar = 1 µm; 12 fps. Temperature increase begins at 1 min, reaching 42°C at ∼5 min. Transgene always speckle-associated: MS2 signal appears without delay (∼1 min after reaching HS temperature at 5 min, see also Fig. 3 A). Transgenes (green), nuclear speckles (lighter green), and MS2-tagged nascent transcripts (red).
