Fig. 7: Inhibitor of Notch signaling pathway tangeretin treatment can improve the lactation function of K14Cre + ; Kindlin-2^flox/flox female mice.

A Therapeutic schedule of tangeretin treatment on K14-Cre + ; Kindlin-2^flox/flox and K14-Cre-; Kindlin-2^flox/flox female mice. Eighteen K14-Cre + ; Kindlin-2^flox/flox female mice weighing 20–25 g were divided into 2 groups at random. The mice caged mating with K14-Cre-; Kindlin-2^flox/flox male mice is recorded as E0. Group 1: nine K14-Cre + ; Kindlin-2^flox/flox female mice were treated with solvent 100 µl from E13.5 and equivalent solvent were used every day. Group 2: nine K14-Cre + ; Kindlin-2^flox/flox female mice were treated with tangeretin 50 mg/kg from E13.5 and equivalent drugs were used every day. Group 3: nine K14-Cre-; Kindlin-2^flox/flox female mice were treated with solvent 100 µl from E13.5 and equivalent solvent were used every day. Group 4: nine K14-Cre-; Kindlin-2^flox/flox female mice were treated with tangeretin 50 mg/kg from E13.5 and equivalent drugs were used every day. The offsprings of these mice was observed and the survival status was recorded. B K-M plot analyses were performed to uncover the difference of survival rate of these offsprings between Tangeretin treatment groups and control groups (Group 1: n = 36, Group 2: n = 40, Group 3: n = 30, Group 4: n = 34). Statistical testing was performed by Log-Rank test. *P < 0.05, **P < 0.01, ***P < 0.001. C Whole-mounted staining of P18.5 mammary gland in K14-Cre + ; Kindlin-2^flox/flox treatment groups or K14-Cre-; Kindlin-2^flox/flox treatment groups. D HE sections staining of P18.5 mammary gland in K14-Cre + ; Kindlin-2^flox/flox treatment groups or K14-Cre-; Kindlin-2^flox/flox treatment groups. Scale bars, 100 µm. E Statistical analysis showing the number of mammary alveolars and cells that make up mammary alveolars in K14-Cre + ; Kindlin-2^flox/flox treatment groups compared with K14-Cre-; Kindlin-2^flox/flox treatment groups (n = 10 fields of view at random per treatment group). Statistical testing was performed by unpaired t test. Data are presented as mean values ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. F Immunohistochemical staining for CK8, Notch1, Hes1 and Hey1 expression of P18.5 mammary gland in four groups. Scale bar, 50 µm. G Quantification for the Notch1 levels in luminal cells (CK8 positive), the target protein of Notch signaling pathway Hes1 and Hey1 levels in the ducts of four groups (n = 6). Statistical testing was performed by unpaired t test. Data are presented as mean values ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.