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. 2022 May 13;12(8):3963–3976. doi: 10.7150/thno.70893

Figure 4.

Figure 4

Gradual initiation of circadian rhythm during hESC (MAN13) chondrogenic differentiation. (A). Successful transduction of clock reporters (hBMAL1-Luc or hPER2-Luc) into cells at day 11 of the differentiation protocol (2D monolayer) or day 21 (10 days in 3D). N = 3, scale bars = 100 µm. (B). At Day 11 (2D), cells were treated with Dex and bioluminescence signals were recorded by the Alligator system. Raw data (Relative light unit, RLU) from a whole well of cells were plotted as intensity over time. Representative traces of 3 independent differentiation runs, with 2-3 technical replicates within each run (N = 3). (C). At Day 21 (3D), pellets transduced with hBMAL1-Luc were synchronised by Dex and bioluminescence signals were recorded by LV200 imaging system. Three independent differentiation runs (N = 3). Raw data (Relative light unit, RLU) from a region of interest encompassing a whole pellet were plotted as intensity over time. 24 hour moving average of the raw data was plotted in the bottom panels.