Journal: Cell Death & Disease

Article Title: CCL2 promotes macrophages-associated chemoresistance via MCPIP1 dual catalytic activities in multiple myeloma

doi: 10.1038/s41419-019-2012-4

Figure Lengend Snippet: a Levels of CCL2, as measured by ELISA, in the peripheral serum of healthy donors (control) and MM patients (MM Pt). Samples from 8 and 15 healthy donors and MM patients, respectively, were used. b The histochemistry score for CCL2 was detected using BM biopsy samples when the patients were newly diagnosed or after treatment (median H-score from 88.104320 to 29.8143). Pt.1 to Pt.12 represent 12 independent patients. c Immunohistochemical analysis of CCL2 expression in 3 representative BM biopsies (Pt.1 to Pt.3). The top panel shows CCL2 expression when patients were newly diagnosed, and the bottom panel shows CCL2 expression after the same patient received therapy. Scale bars, 50 μm. d Ratio of mRNA expression of CCL2 in MM cell lines (RPMI 8226, MM.1 S, CAG, JJN3, OPM2, ARP-1), PBMCs and macrophages (Mφs) by RT-PCR. e A total of 1 × 10 5 cells were cultured in 1 mL of medium for 24 h, followed by ELISA analysis of CCL2 expression in cell culture supernatants. f, g Mφs were cultured alone (Mφs) or cocultured with MM cells directly (Mφs + ARP-1, Mφs + MM.1S) or through Transwell chambers (Mφs/ARP-1, Mφs/MM.1S) for 24 h. Then, the fresh median was changed, and the Mφs were cultured for another 24 h, followed by RT-PCR f and ELISA g to detect CCL2 expression. Summarized data from at least three independent experiments are shown. Values are presented as means ± SD. * P < 0.05; ** P < 0.01; *** P < 0.001

Article Snippet: A neutralizing anti-CCL2 antibody (αCCL2, 50 μg/ml) was purchased from R&D Systems, MN, USA.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Immunohistochemical staining, Expressing, Reverse Transcription Polymerase Chain Reaction, Cell Culture

Journal: Cell Death & Disease

Article Title: CCL2 promotes macrophages-associated chemoresistance via MCPIP1 dual catalytic activities in multiple myeloma

doi: 10.1038/s41419-019-2012-4

Figure Lengend Snippet: a Mφs generated from PBMCs of two independent healthy donors (Mφs-1 and Mφs-2) were treated with rhCCL2 (50 ng/ml) for 24 h. RNA was extracted for RNA sequencing. Heat maps illustrate the log 2 -fold change of some immune-related genes. b Mφs were exposed to rhCCL2 (50 ng/mL) for the indicated times, followed by RT-PCR and Western blotting c to determine expression of MCPIP1. d Mφs were treated with rhCCL2 (50 ng/mL) for 24 h. Immunofluorescence was performed to detect expression of MCPIP1 in Mφs. Scale bar, 50 μm. e , f Mφs were treated with RPMI-8226 CM for 24 h with or without αCCL2. qRT-PCR e and Western blotting f were performed to detect MCPIP1 expression. Data are presented as means ± SD of at least three independent experiments. * P < 0.05; ** P < 0.01, *** P < 0.001

Article Snippet: A neutralizing anti-CCL2 antibody (αCCL2, 50 μg/ml) was purchased from R&D Systems, MN, USA.

Techniques: Generated, RNA Sequencing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing, Immunofluorescence, Quantitative RT-PCR

Journal: Cell Death & Disease

Article Title: CCL2 promotes macrophages-associated chemoresistance via MCPIP1 dual catalytic activities in multiple myeloma

doi: 10.1038/s41419-019-2012-4

Figure Lengend Snippet: CCL2 strongly recruits monocytes to the BM microenvironment. Then these recruited Mφs interact with MM cells and further upregulated Mφs’ expression of CCL2. These increased CCL2-induced MCPIP1 expression in Mφs through STAT3 pathway, polarizing Mφs toward M2-like phenotype and promoting Mφs to protect MM cells from drug-induced apoptosis

Article Snippet: A neutralizing anti-CCL2 antibody (αCCL2, 50 μg/ml) was purchased from R&D Systems, MN, USA.

Techniques: Expressing