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

Figure 5.

Figure 5

Ectopic persistent HES1 expression does not increase total HES1 level above physiological range

(A) Example pictures of E13.5 HES1:mSCARLET-I NSCs transfected with UbC-mVENUS:HES1 reporter. White arrows mark transfected (top panels) or untransfected (bottom panels) cells where FCS was performed (1). Schematic showing how total HES1 concentration is compared between transfected and untransfected cells. Green dots represent UbC-mVENUS:HES1 concentration and red dots represent HES1-mSCARLET-I concentration (2) (scale bars = 5μm).

(B and C) Box plots showing total HES1 concentration in untransfected and UbC-mVENUS:HES1 transfected E13.5 HOM HES1:mSCARLET-I NSCs. In the reporter transfected cells, the total HES1 concentration is the sum of UbC-mVENUS:HES1 concentration and HES1:mSCARLET-I concentration. In (B) all concentrations have been estimated experimentally by FCS (apart from the HES1:mSCARLET-I concentration in transfected cells which was inferred). In (C) all concentrations have been inferred (black horizontal lines represent median, dots represent individual cells, number of cells analyzed in B) untransfected= 75, transfected = 38, n = 3 biological replicates, in (C) untransfected = 316, transfected = 77, n = 3 biological experiments, two-tailed paired ttest, ∗p = 0.047, ∗∗p = 0.008).

(D) Bar graph showing the percentage of E13.5 HOM HES1:mSCARLET-I NSCs transfected with UbC-mVENUS:HES1 that have a total HES1 concentration (either experimentally estimated or inferred) within the range of total HES1 concentration values in their untransfected counterparts.