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. 2022 Jan 11;11(2):229. doi: 10.3390/cells11020229

Figure 4.

Figure 4

The MyD88–JAK2/TYK2–STAT-3–Sec22b pathway is involved in the repression of macrophage phagocytosis by eHSP90α. (A) Phagocytotic activities of the THP-1-derived macrophages treated for 24 h with PBS, rHSP90α, or rHSP90α plus control IgG or anti-CD91 or TLR4 antibody. The quantitative data are the mean ± SD of three independent experiments. a p < 0.01 when compared with “PBS” group. b p < 0.01 when compared with “rHSP90α + IgG” group. (B) Phagocytotic activities of the THP-1-derived macrophages treated with PBS or rHSP90α plus DMSO, JSI-124, or JAKi. The quantitative data are the mean ± SD of three independent experiments. # p < 0.01 when compared with “rHSP90α + DMSO” group. (C) mRNA levels of Sec22b and Syntaxin 18 in the THP-1-derived macrophages treated for 24 h with PBS or 15 μg/mL of rHSP90α. The representative RT-PCR data of three independent experiments are shown. (D) Phagocytotic activities of the Sec22b-knockdown macrophages treated with PBS or rHSP90α. Two THP-1 cell clones expressing Sec22b-targeting shRNA #1 and #2, respectively, were induced by 12-O-tetradecanoyl-13-phorbol acetate to differentiate into macrophages. The macrophages were then treated with PBS or rHSP90α for 24 h. The quantitative data are the mean ± SD of three independent experiments. @ p < 0.01 when compared with “–” group. (E) Sec22b mRNA levels in the THP-1-derived macrophages treated for 24 h with PBS or rHSP90α plus DMSO, JSI-124, or JAKi. The RT-PCR data shown are the representative of three independent experiments. (F) ChIP assay showing rHSP90α-induced binding of STAT-3 to sec22b gene promoter. The representative data of three independent experiments is shown.