Figure 3.

Regulatory T-cell recruitment to HNSCC tumor sites is inhibited by dietary black raspberry extract administration. (A) Multiplex confocal microscopy images of representative tongues taken from each mouse group. For each image, individual stains and composite images are shown. Each tongue was stained for DAPI (blue), CD4 (red), and FoxP3 (green). FoxP3⁺ CD4⁺ cells were quantified using ImageJ. (B) Western blot demonstrating FoxP3, and GAPDH protein content of tongue lysates isolated from sentinel, 4NQO, and 4NQO+BRB-E groups. Average protein intensity of Western blot bands was measured for several samples using ImageJ. (C) RT-qPCR data of tongue homogenate taken from sentinel or cancer-bearing mice receiving the control diet or the black raspberry extract supplemented feed for Foxp3, Il2r, and Ctla4. Data are represented as fold change relative to the Bactin control. *p-value < 0.05; **p-value < 0.01; ***p-value < 0.001; #p-value < 0.1 for comparisons of BRB-E treatment groups to the 4NQO-exposed control diet group using Student’s t-test.