Expression of lnc-C/EBPβ in inflammation and in MDSCs induced by tumor-associated factors. Expression of lnc-C/EBPβ in inflammation and in MDSCs induced.

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Expression of lnc-C/EBPβ in inflammation and in MDSCs induced by tumor-associated factors. Expression of lnc-C/EBPβ in inflammation and in MDSCs induced by tumor-associated factors. A, Schematic diagram showing differentiation of MDSCs. B, LncRNA microarray of MDSCs. MDSCs were exposed to GM-CSF or GM-CSF plus IL6 (GM + IL6) for 24 hours. C, QRT-PCR (top) and RT-PCR (bottom) of lnc-C/EBPβ in MDSCs. U6 RNA was used as an internal control. GM: GM-CSF; GM/IL6: GM-CSF plus IL6. D, Homologue analyses of lnc-C/EBPβ through mVista and rankVista. Blue: lnc-C/EBPβ sequence; peaks over dark line: conserved regions. E, RT-PCR (left) and northern blot (right) of murine lnc-C/EBPβ in mouse MDSCs. F, RT-PCR (left) and northern blot (right) of homologous lnc-C/EBPβ in human monocytes. G, Fluorescence in situ hybridization of lnc-C/EBPβ in mouse MDSCs (M-MDSC) and human monocytes (Hu-monocyte). NC-FAM: control probe; Lnc-C/EBPβ-FAM: FAM-labeled lnc-C/EBPE probe. H, qRT-PCR of mouse lnc-C/EBPβ (mlnc-C/EBPβ) in MDSCs after exposure to different cytokines and tumor supernatants. GM: GM-CSF; 1D8: mouse ovarian cancer cell line; 4T1: mouse breast cell line; B16: mouse melanoma cell line; Lewis, mouse lung cancer. I, QRT-PCR of human lnc-C/EBPβ (hulnc-C/EBPβ) in human MDSC-like cells after exposure to different cytokines and tumor supernatants. GM: GM-CSF; MCF: human breast cancer cell line; HeLa: human cervical cancer cell line; HT29: human colon cancer cell line; MDA: human breast cancer cell line. J, qRT-PCR of lnc-C/EBPβ in MDSCs isolated from the spleens of mice bearing breast cancer 4T1, melanoma B16, or ovarian cancer 1D8. K, QRT-PCR of mouse lnc-C/EBPβ in JAK1 and STAT3 inhibitor–treated MDSCs. L, qRT-PCR of lnc-C/EBPβ in MDSCs isolated from the spleens of wild-type (WT) and IL6−/− mice bearing B16 tumors. M, QRT-PCR of mouse lnc-C/EBPβ in the different immune cells from the spleens of mice bearing melanoma B16. Two-tailed, paired t test was used in C, H–M. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Also see Supplementary Figs. S1–S3, and GSE104718, as well as GSE104719. Data are representative of three independent experiments. Yunhuan Gao et al. Cancer Immunol Res 2018;6:1352-1363 ©2018 by American Association for Cancer Research