Fig. 6.

CPEB3 attenuates tumorigenesis and TAM polarization in vivo (a) Schematic of the procedure for separating tumor cells and TAMs. (b) HCT116 cells were stably infected with Ctrl and CPEB3 lentivirus, and LoVo cells were stably infected with shCtrl and shCPEB3 sequences. Tumorigenesis assay of Balb/c nude mice subcutaneously injected with HCT116-Ctrl/CPEB3 cells and LoVo-shCtrl/shCPEB3 cells (n = 20). Representative photos of tumors from mice in various groups. (c) IHC staining of Ki67 positive cells was counted per high-power field (PHF), while E-cadherin and vimentin expression scores were counted in tumor tissues in a mouse xenograft model; error bars, SEM. (d) The mice with intra-spleen injection of LoVo-shCtrl/shCPEB3 cells were treated with tocilizumab (5 mg/kg) weekly via intraperitoneally injection. The number of liver metastatic sites (indicated by arrows) was counted under the microscope; error bars, SEM. (e) Macrophages were separated from murine tumor tissues using Percoll-layered liquid. Surface expression of CD86 and CD163 was detected in macrophages using flow cytometry. The percentage of CD86⁺ or CD163⁺ cells in macrophages was reported using error bars and SEM. (f) Expression of JAK1, pJAK1, STAT3, and pSTAT3 in the tumor tissues of the two groups were analyzed by western blot analysis. (g) Schematic overview of the mechanisms by which CPEB3 modulate TAM polarization and inhibit colorectal cancer EMT. ** P < 0.01; *** P < 0.001; **** P < 0.0001