Figure 4.

Rac1 potentiates primary HBC differentiation in vitro

(A–D) Representative immunofluorescence images of Hopx expression in cultures of vehicle-treated (A), activated (B), activated and Rac1 inhibited (C), and Rac1 inhibited (D) HBCs.

(E) Quantification of norm. total Hopx fluor. density for conditions represented in (A) (4,927 CK5⁺ HBCs), (B) (8,065 CK5⁺ HBCs), (C) (7,235 CK5⁺ HBCs), and (D) (5,358 CK5⁺ HBCs) (n = 4 independent trials), each circle represents an analyzed CK5⁺ HBC.

(F–I) Representative immunofluorescence images of Ki67 expression in cultures of vehicle-treated (F), activated (G), activated and Rac1 inhibited (H), and Rac1 inhibited (I) HBCs.

(J) Quantification of norm. nuclear Ki67 fluor. density for conditions represented in (F) (4,716 CK5⁺ HBCs), (G) (7,325 CK5⁺ HBCs), (H) (7,297 CK5⁺ HBCs), and (I) (5,228 CK5⁺ HBCs) (n = 4 independent trials); each circle represents an analyzed DAPI⁺ area within a CK5⁺ HBC. Vehicle (PMA⁻/EHT1864^-) used as baseline; error bars indicate mean ± SEM, Kruskal-Wallis test with post hoc Dunn’s multiple comparisons test, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 (E, J). Scale bar equals 50 μm (A).