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. 2022 Sep 2;28(11):1733–1747. doi: 10.1111/cns.13905

FIGURE 6.

FIGURE 6

NEDD4L downregulates PIK3CA to inhibit the PI3K/AKT pathway, thus restraining M2 polarization of macrophages and glioma cell proliferation, migration, and invasion. (A) The protein expression of NEDD4L, PIK3CA, and AKT and phosphorylated AKT in U937 cells after overexpression NEDD4L and PIK3CA examined by Immunoblotting. (B) The mRNA expression of iNOS, TNF‐α, Arg‐1, and IL‐10 in U937 cells after overexpression NEDD4L and PIK3CA measured by RT‐qPCR. (C) Immunoblotting of protein expression of iNOS and Arg‐1 in U937 cells after overexpression NEDD4L and PIK3CA. (D) The expression of TNF‐α and IL‐10 in supernatant of U937 cells after overexpression NEDD4L and PIK3CA assessed by ELISA. (E) The proportion of CD11b+CD163+ cells in U937 cells after overexpression NEDD4L and PIK3CA evaluated by flow cytometry. (F) The proliferation in U87 and A172 cells co‐cultivated with U937 cells tested by EdU. (G) Cell migration in U87 and A172 cells co‐cultivated with U937 cells examined by Transwell. (H) Cell invasion in U87 and A172 cells co‐cultivated with U937 cells examined by Transwell. *p < 0.05 versus U937 cells transfected with oe‐NC or U87 or A172 co‐cultivated with U937 cells treated without anything. # p < 0.05 versus U937 cells transfected with oe‐PIK3CA or U87 or A172 co‐cultivated with U937 cells transfected with oe‐PIK3CA. The experiment was repeated three times