Fig. 9.

Elevated frequencies of CD45⁺ cells were observed in tumors treated with either nanoformulated CCL21 or CCL21 in comparison to the control. Neuro2a tumor-bearing mice received 2 daily injections (1 in the morning and 1 in the evening) for 2 days in a row with each injection containing 6 μg of CCL21/25 μL, or 6 μg of CCL21 in nanoformulation/25 μL per dose, or an equal volume (25 μL) of buffer. The frequency of CD45⁺ cells was significantly (p < 0.0001) higher in tumors treated with either nanoformulated CCL21 or CCL21 compared to those treated with buffer control at day 2 post treatment initiation. Significantly higher percentages of CD45⁺ cells were also seen at day 3 post treatment in nanoformulated CCL21-treated tumors (p = 0.0190) or CCL21-treated tumors (p = 0.0090) compared to the control. Gating strategy for CD45⁺ cells: single cells, live cells, CD45⁺. Frequency was calculated as % marker = (number of cells that were marker positive) / (number of live cells) X 100. Graphical representation of the data depicts the mean frequency overlaid with individual data points for each group. Statistical comparisons were made via two-way ANOVA. The bars indicate standard deviation. Statistical significance was defined as p < 0.05*, p < 0.01**, p < 0.001***, and p < 0.0001****. Groups consisted of 5–10 mice per treatment group per time point.