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. 2021 Oct 2;24(10):103198. doi: 10.1016/j.isci.2021.103198

Figure 1.

Figure 1

Characteristics of HES1 dynamic expression in proliferative, quiescent and reactivated conditions

(A) Schematic structure of LUC2:HES1 BAC used to generate transgenic mice. NSC lines were established from the LGE region of E13.5 LUC2:HES1 embryos (CTX = cortex, LGE = lateral ganglionic eminence, MGE = medial ganglionic eminence).

(B) Experimental design for induction of quiescence and reactivation (upper panel). Example images from the analysis of cell proliferation by Ki67 staining at proliferative, quiescent and reactivated conditions. Cells were counterstained with DAPI (bottom panels) (scale bars = 30μm).

(C) Percentage of Ki67 positive cells in proliferative, quiescent and reactivated conditions (error bars represent standard deviation, n = 9 biological experiments, One-way ANOVA with Uncorrected Dunn’s multiple comparison test, proliferative versus quiescent ∗∗∗∗p < 0.0001, quiescent versus reactivated ∗p = 0.04, proliferative versus reactivated ∗p = 0.04).

(D) Example cell traces of E13.5 LUC2:HES1 NSCs cultured in proliferative, quiescent and reactivated conditions (A.U. = arbitrary units).

(E) Percentage of oscillators in proliferative, quiescent and reactivated conditions (error bars represent standard deviation, number of 10-h cell traces analyzed: 75 proliferative from n = 8, 171 quiescent from n = 4, 182 reactivated from n = 5 biological experiments).

(F–H) Box plots representing analysis of 10h bioluminescence traces for period of oscillations (F), quality of oscillations (G) and maximum peak-to-trough ratio per oscillatory trace (H). Note that the first 10h of imaging of individual cells have a random starting point from the start of the imaging of the culture (dots represent individual oscillatory cell traces, black horizontal lines represent median, number of 10-h oscillatory cell traces analyzed: 56 proliferative from n = 7, 110 quiescent from n = 4, 149 reactivated from n = 5 biological experiments, Kruskal-Wallis test with Dunn’s multiple comparison test, ∗p = 0.038, ∗∗∗p = 0.001, ns = not significant)