Fig. 3.

EMP3 induces CCL2 and TGF-β1 in GBM cells to promote M2 TAM recruitment and polarization. a CCL2 and TGF-β1 mRNA expression in EMP3_KO and EMP3_Scra GL261 cells was measured by qRT-PCR. The expression of these transcripts was normalized to that of β-actin. Student’s t-test was performed. KO: Knockout; Scra: Scramble. b The expression of CCL2 and TGF-β1 in EMP3_KO and EMP3_Scra GBM cell supernatants was measured by ELISA. Student’s t-test was performed. KO: Knockout; Scra: Scramble. c-d The ARG-1, MRC1, IL-10, and TNF-α mRNA expression in RAW264.7 cells exposed to supernatants from EMP3_OE, EMP3_Scra, EMP3_OE+anti-TGF-β1/anti-CCL2, or EMP3_OE+IgG GL261 cells for 48 hours was detected by qRT-PCR. The expression of these transcripts was normalized to that of β-actin. Student’s t-test was performed. OE: Overexpression; Scra: Scramble. e-f Migration assays performed with RAW264.7 cells exposed to supernatants from EMP3_OE, EMP3_Scra, EMP3_OE+anti-TGF-β1/anti-CCL2, or EMP3_OE+IgG GL261 cells for 48 hours. Scale bar = 100 μm. OE: Overexpression; Scra: Scramble. Student’s t-test was performed.g-h Representative images of IF staining for CD86 and CD206 in BV-2 cells exposed to supernatants from EMP3_OE, EMP3_Scra, EMP3_OE+anti-TGF-β1/anti-CCL2, or EMP3_OE+IgG GL261 cells for 48 hours. The histogram summarizes the relative fluorescence intensity of the cells. Scale bar = 100 μm. Student’s t-test was performed. KO: Knockout; Scra: scramble. The mean ± S.D. is shown. Ns: nonsignificant, *p < 0.05, **p < 0.01, and ***p < 0.001