Journal: Cancers

Article Title: Cancer-Associated Fibroblasts Exposed to High-Dose Ionizing Radiation Promote M2 Polarization of Macrophages, Which Induce Radiosensitivity in Cervical Cancer

doi: 10.3390/cancers15051620

Figure Lengend Snippet: CCL2 is critical for high-dose irradiated CAFs to promote M2 polarization. ( A ) mCAFs were exposed to 8 Gy radiation or not (control), and the supernatant of mCAFs was subjected to multiplex cytokine array analysis 24 h later. Left: a representative blot. Right: quantification of dot intensity of significantly changed cytokines. ( B ) Cervical cancer cell lines and CAFs were irradiated with the indicated doses. After 24 h, CCL2 expression was analyzed by Western blot analysis. ( C ) mCAFs and hCAFs were exposed to the indicated doses of radiation. After 24 h, the supernatant was subjected to ELISA. Data are shown as mean ± SD from three independent experiments. The data were analyzed using one-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001. ( D ) BMDMs were treated with 20 ng/mL CCL2 for 24 h and then subjected to flow cytometry analysis. Data are shown as mean ± SD from three independent experiments. Differences between groups were analyzed using unpaired Student’s t -test. * p < 0.05. ( E ) M0 BMDMs treated with 10 nM CCR2 antagonist (INCB3344) were co-cultured with irradiated or non-irradiated mCAFs for 3 days. Then, the cells were subjected to flow cytometry analysis to evaluate CD206 expression. Data are shown as mean ± SD from three independent experiments. * p < 0.05. ( F ) Anti-CCL2 neutralizing antibody (20 μg/mL) was added to a co-culture system consisting of 8 Gy irradiated mCAFs and M0 BMDMs. The BMDMs were subjected to flow cytometry analysis after 3 days of co-cultivation. Data are shown as mean ± SD from three independent experiments. The data were analyzed using one-way ANOVA. * p < 0.05. ( G ) M0 BMDMs were co-cultured with mCAFs exposed to the indicated doses of radiation for 3 days and 10 nM CCR2 antagonist (INCB3344) was added into the 8 Gy irradiated mCAF co-culture system. Immunofluorescence analysis of the expression of Ym-1 (red) in BMDMs. The whole western blots are shown in File S1.

Article Snippet: The supernatant of CAFs and cervical cancer cells was assayed for cytokines and chemokines using the Mouse XL Cytokine Array (Ary028; R&D Systems) and CCL2 ELISA kits (mouse, EK0568; human, EK0441; BOSTER) according to the manufacturers’ instructions.

Techniques: Irradiation, Control, Multiplex Assay, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Cell Culture, Co-Culture Assay, Immunofluorescence

Journal: Cancers

Article Title: Cancer-Associated Fibroblasts Exposed to High-Dose Ionizing Radiation Promote M2 Polarization of Macrophages, Which Induce Radiosensitivity in Cervical Cancer

doi: 10.3390/cancers15051620

Figure Lengend Snippet: Model summarizing the proposed signaling events among CAFs, TAMs, and cervical cancer cells under high-dose radiation. CAFs secrete an elevated level of CCL2 upon treatment with high-dose radiation. CCL2 promotes pro-tumor transition in macrophages. M2 macrophages induce radioresistance in cervical cancer cells.

Article Snippet: The supernatant of CAFs and cervical cancer cells was assayed for cytokines and chemokines using the Mouse XL Cytokine Array (Ary028; R&D Systems) and CCL2 ELISA kits (mouse, EK0568; human, EK0441; BOSTER) according to the manufacturers’ instructions.

Techniques:

Journal: Cell metabolism

Article Title: Low-Dose Sorafenib Acts as a Mitochondrial Uncoupler and Ameliorates Nonalcoholic Steatohepatitis

doi: 10.1016/j.cmet.2020.04.011

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse MCP1(CCL2) ELISA kit , Cloud-Clone Co. , Cat#SEA087Mu.

Techniques: Recombinant, Protease Inhibitor, Western Blot, SYBR Green Assay, Modification, Bicinchoninic Acid Protein Assay, Cholesterol Assay, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Gene Knockout, Knock-Out, Plasmid Preparation, Software

Journal: Cell metabolism

Article Title: Low-Dose Sorafenib Acts as a Mitochondrial Uncoupler and Ameliorates Nonalcoholic Steatohepatitis

doi: 10.1016/j.cmet.2020.04.011

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse MCP1(CCL2) ELISA kit , Cloud-Clone Co. , Cat#SEA087Mu.

Techniques: Recombinant, Protease Inhibitor, Western Blot, SYBR Green Assay, Modification, Bicinchoninic Acid Protein Assay, Cholesterol Assay, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Gene Knockout, Knock-Out, Plasmid Preparation, Software