Reduced in vivo Notch signaling results in reduced Foxp3 expression within the Treg pool and spontaneous lymphocyte infiltration of the liver. For 3 months, wild-type mice were fed either normal rodent chow (control, n = 4) or chow formulated with LY 411575 (GSI, n = 5) to deliver 5 mg/kg per day. (A) Bulk splenocytes were stained for flow cytometry with antibodies specific for CD4 and Foxp3. Values represent mean percentages. (B) Histogram representing level of Foxp3 expression, gated on live CD4+Foxp3+ cells. Data are representative of data collected from 2 independent experimental groups (group 1: control, n = 2 and GSI, n = 3; group 2: control, n = 2 and GSI, n = 2). (C) Graphic representation of mean fluorescence intensity of Foxp3 staining in all experimental groups from panel B. Values represent the mean. (D-G) H&E staining of livers from mice with normal (D,F) or reduced (E,G) levels of Notch1 signaling. One representative liver section from each group is shown: (D) control mouse, (E) GSI chow-fed mouse, (F) wild-type mouse (n = 6), (G) Notch1 antisense mouse (n = 6). A Spot digital camera (Diagnostic Instruments, Sterling Heights, MI) mounted on a Zeiss Axioscope microscope (Carl Zeiss, Jena, Germany) using a 20× objective was used to acquire figures.
