Fig. 3

Effects of lncGBP9 on macrophage polarization in vitro. a BMDMs were induced toward M0 macrophages for 7 days as described in the M&M section. Then, the silencing and overexpression of lncGBP9 conducted in M0 macrophages for 48 h, as confirmed by real-time PCR. b, c Adv-sh-lncGBP9 or Adv-lncGBP9 transduced M0 macrophages were induced toward M1 polarization for 48 h and the mRNA expression and protein levels of STAT1, p-STAT1, and SOCS3 in response to lncGBP9 silencing or lncGBP9 overexpression were determined by real-time PCR and Immunoblotting in M1 macrophages. d The production of IL-6 and IL-12 in response to lncGBP9 silencing or lncGBP9 overexpression was determined by ELISA in M1 macrophages. e, f Adv-lncGBP9 or Adv-sh-lncGBP9 transduced M0 macrophages were induced toward M2 polarization for 48 h and the mRNA expression and protein levels of STAT6, p-STAT6, and SOCS3 in response to lncGBP9 overexpression or lncGBP9 silencing were determined by real-time PCR and Immunoblotting in M2 macrophages. g The production of IL-10 and TGF-β1 in response to lncGBP9 overexpression or lncGBP9 silencing was determined by ELISA in M2 macrophages. Next, M1 macrophages were co-transfected with Adv-lncGBP9 and SOCS3-overexpressing vector (SOCS3 OE) and examined for h the mRNA of STAT1 and SOCS3 by real-time PCR. i The protein levels of SOCS3, STAT1, and p-STAT1 by Immunoblotting. j The concentrations of IL-6 and IL-12 by ELISA. M2 macrophages were co-transfected with Adv-lncGBP9 and si-SOCS3 and examined for k the mRNA of STAT6 and SOCS3 by real-time PCR. l The protein levels of SOCS3, STAT6, and p-STAT6 by immunoblotting. m The concentrations of IL-10 and TGFβ1 by ELISA. *P < 0.05, **P < 0.01, compared to the control group; #P < 0.05, ##P < 0.01, compared to the Adv-lncGBP9 + NC (negative control) vector or Adv-lncGBP9 + si-NC (negative control) group. Values are mean ± S.D of n = 3 independent experiments