A Novel In Situ Dendritic Cell Vaccine Triggered by Rose Bengal Enhances Adaptive Antitumour Immunity

Figure 3

Lewis cancer stem cells treated with RB were exposed to calreticulin on the surface (ecto-CRT). (a) The increased size of the cell spheres after 7 days (magnification of 200x). (b) The results of the real-time quantitative PCR analysis indicated that the expression of the Oct-4, CD44, and ABCG2 genes was significantly elevated in Lewis CSCs than LLCs. (c–e) The RB treatment induced increased ecto-CRT expression in CSCs in a dose-dependent manner. Increased expression was observed on both live (d) and dead cells (e) based on flow cytometric analysis. The gating strategy of ecto-CRT expression in cancer stem cells: First, the cells were collected, and the voltage was adjusted according to the blank control. Then, a new scatterplot was created, and the target cells were gated (cross gate) and finally compensated by APC, PE, FITC, and DAPI single-stained controls. A representative example of three separate experiments is summarized in (d). The are shown. One-way ANOVA was performed, and the significance level was defined as , , and .

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