Journal: iScience
Article Title: Targeting USP14 enhances immunotherapy response by reprogramming tumor-associated macrophages in colon cancer
doi: 10.1016/j.isci.2026.115362
Figure Lengend Snippet: Targeting USP14 inhibits tumor growth and induces local anti-tumor immunity in vivo (A) Representative images of MC38 tumors harvested on day 21 post-inoculation from mice treated with IU1 (20 mg/kg, i.p., on days 6, 9, 12, and 15) or vehicle control. (B) Statistics of MC38 tumor growth rate following IU1 or vehicle treatment in vivo . The intraperitoneal injection dose of IU1 was 20 mg/kg, administered on days 6, 9, 12, and 15. Data are presented as the mean ± SEM ( n = 6 per group). (C) Spider diagram of the tumor volume growth in each mouse from the IU1 group and the PBS group. (D) Gating strategy for the detection of the TAMs by flow cytometry. We first obtained live cells, and then identified cells that were positive for CD45, CD11b, F4-80, and CD206 as M2 macrophages. (E–P) Proportions of neutrophil (E), M2 macrophage (F), M1 macrophage (G), MDSC (H), activated DCs (I), CD4 T cell (J), Treg cells (K), CD8 T cell (L), IFN-γ + CD8 T cell (M), precursor exhausted T cells (TCF-1 + ) (N), effective CD8 T cells (PD-1 + ) (O) and activated CD8 T cell (CD69 + ) (P) in the TME of the IU1 group and the control group by using flow cytometry. (Q–U) Cytokines IFN-γ (Q), TNF-α (R), IL-2 (S), IL-10 (T), and IL-12 (U) in the TME of each group were detected by Mul-Analyte Flow Assay Kit. (V) Schematic illustration of the proposed mechanism of action of IU1 in reprogramming the tumor microenvironment. Data are presented as the mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001, and ns: not significant.
Article Snippet: For RNA and protein extraction, CD11b + cells were first enriched using magnetic-activated cell sorting (MACS) with anti-CD11b microbeads (Miltenyi Biotec) according to the manufacturer’s protocol.
Techniques: In Vivo, Control, Injection, Flow Cytometry