Fig 1: CGF extract regulated the macrophage-mediated immune response via the AKT pathway. (A) A western blot analysis of the proteins involved in the PI3K/AKT (p-AKT, AKT and PI3K) and JAK/STAT (p-JAK, JAK, p-STAT3 and STAT3) signaling pathways in CCM-treated THP-1 macrophages. AKT phosphorylation was increased, but no significant differences were observed in the JAK/STAT pathway (n = 3). (B) The effect of an AKT inhibitor on AKT pathway inhibition was analyzed using western blotting (n = 3). (C) Representative images of adherent THP-1 macrophages cultured in CCM with or without AKT inhibitor were analyzed using ImageJ software (n = 5). (D) The secretion of IL-1ß and RANTES by THP-1 macrophages were assayed using ELISAs (n = 3). (E) Expression of the M2 subtype marker CD163 was detected using immunofluorescence staining (n = 3). The data are presented as the mean ± SD and were analyzed using one-way analysis of variance (ANOVA) and post hoc tests: *P < 0.05, **P < 0.01 and ***P < 0.001.
Fig 2: CGF extract modulated macrophage cytokine secretion. (A) The supernatants of CCM-treated macrophages were screened using a human cytokine antibody array. (B) Representative cytokines are presented. (C) A GO analysis of differentially expressed cytokines enriched in BP terms after 20% CCM treatment. (D) Enriched MF categories. (E) The mRNA expression of differentially expressed cytokines (IL-1ß, IL-7, RANTES and MCP-1) in CCM-treated macrophages was analyzed using q-PCR. (F) The secretion of IL-1ß and RANTES by CCM-treated macrophages was further confirmed using ELISAs. The quantitative data are presented as the mean ± SD (n = 3) and were analyzed using one-way analysis of variance (ANOVA) and post hoc Dunnett tests: *P < 0.05, **P < 0.01 and ***P < 0.001.
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