Fig. 6

Lactate-stimulation GPR65 on TAMs promoted glioma cells malignant progression via secreting HMGB1. A RT-PCR analysis of various potential TAM-derived chemokines and cytokines in PMA-treated THP1 cells transfected with shNC or shGPR65, with or without lactate stimulation. B RT-PCR and (C) Western blot analysis of HMGB1 expression in macrophages under lactate concentration gradient stimulations. D HMGB1 concentrations in CMs from macrophages under lactate concentration gradient stimulations. E Western blot analysis of HMGB1 expression in macrophages transfected with shNC or shGPR65, with or without lactate stimulation. F HMGB1 concentrations in CMs from macrophages transfected with shNC or shGPR65, with or without lactate stimulation. G-H CCK8 (G) and clone formation assays (H) were performed to assess cell proliferation of U87 and U251 cells stimulated with CMs from various pretreated macrophages with HMGB1 inhibitor or rHMGB1. I-J Wound healing (I) and transwell assays (J) were conducted to evaluate cell migration and invasion of U87 and U251 cells stimulated with CMs from various pretreated macrophages with HMGB1 inhibitor or rHMGB1. K Western blot analysis of Vimentin, N-cadherin, and E-cadherin protein expression levels of U87 and U251 cells stimulated with CMs from various pretreated macrophages with HMGB1 inhibitor or rHMGB1. Data is presented as the Mean ± SEM. *p < 0.05, **p < 0.01, and ***p < 0.001 vs. shNC group. ^nsp > 0.05, and ^###p < 0.001 vs. shGPR65 group