Fig. 1.

HBCs at 24 hpi exhibit dynamic Notch pathway protein expression that occurs concomitant with their activation. (A-D) Relative to their counterparts within uninjured (uninj.) OE (A,C), HBCs at 24 hpi (B,D) are activated, given the decrease in nuclear p63. Each CK5⁺ Nuclear Object (Obj.) is derived from a DAPI⁺ nucleus within a CK5⁺ HBC. (E) Quantification of HBC nuclear p63 normalized (norm.) fluorescence (fluor.) density, each point represents a CK5⁺ Nuclear Object (as represented by blue outlines in C and D) (n=3 mice, 3829 uninjured HBCs and 1419 24 hpi HBCs). (F-I) Total HBC-expressed Notch1 (within magenta outlines, H and I) is unchanged at 24 hpi. (J) Quantification of HBC total Notch1 normalized fluorescence density, each point represents a CK5 Object (as represented by magenta outlines in H and I) (n=3 mice, 2833 uninjured HBCs and 1126 24 hpi HBCs). (K-N) Relative to their counterparts within uninjured OE (K,M), HBCs at 24 hpi (L,N) demonstrate an increase in nuclear Hes1. Each CK5⁺ Nuclear Object is derived from a DAPI⁺ nucleus within a CK5⁺ HBC. (O) Quantification of HBC nuclear Hes1 normalized fluorescence density, each point represents a CK5⁺ Nuclear Object (as represented by blue outlines in M and N) (n=3 mice, 2201 uninjured HBCs and 983 24 hpi HBCs). (P-S) Total HBC-expressed Dll1 (within magenta outlines, R and S) is significantly decreased at 24 hpi. (T) Quantification of HBC total Dll1 normalized fluorescence density, each point represents a CK5 Object (as represented by magenta outlines in R and S) (n=3 mice, 4739 uninjured HBCs and 2688 24 hpi HBCs). Uninjured was used as baseline, data are mean±s.e.m., Mann–Whitney test, ****P<0.0001 (E,J,O,T). Scale bars: 20 μm.