Fig. 2.

LincRNA-p21 knockdown reversed the functional phenotype of TAMs in tumor milieu. a The transfection efficiency reached 96% (left) and the relative expression of lincRNA-p21 (right). Fluorescent labeled siRNAs were designed to down-regulate lincRNA-p21 in macrophages. The transfection efficiency was observed under fluorescence microscope after 6 h, lincRNA-p21 expression was detected by RT-qPCR after 24 h. Data are presented as mean ± SEM. b CD86 and MHC II up-regulated; conversely, CD206 down-regulated following lincRNA-p21 knockdown in TAMs. The upper panels showed representative images and lower panels showed statistical analysis. c LincRNA-p21 knockdown promoted pro-inflammatory cytokines’ secretion of TAMs. Secretion of IL-4, IL-6, IL-10, IL-12 and TNF-α was assessed by ELISA in the supernatant. d The levels of iNOS up-regulated; however, Arg-1 down-regulated following lincRNA-p21 knockdown in TAMs. Western blotting analysis was employed to assess the levels of iNOS and Arg-1. β-actin was used as a loading control. The left panels show representative blots and right panels show statistical analysis. e LincRNA-p21 knockdown could not increase phagocytosis of TAMs. LincRNA-p21 was knockdown in TAMs, then Escherichia coli (E. coli) was added. After 1 h, the bacteria were sterilized, TAMs were lysed. The number of bacteria phagocytized by TAMs was counted. The left panels show representative images and right panels show statistical analysis. Data are presented as mean ± SEM. All the data were obtained from three independent experiments and the representative images are shown. *P < 0.05 relative to control. N.S. means no significance. SLP1 means lincRNA-p21 downregulated TAMs