Journal: PLOS One
Article Title: JMJD3 regulates the M2-like macrophage polarization and promotes the growth of breast cancer cells via STAT6/IRF4 axis
doi: 10.1371/journal.pone.0341313
Figure Lengend Snippet: (A) THP-1 cells were infected with shJMJD3 or oeJMJD3 lentiviruses for 72 hours. JMJD3 mRNA expression was then assessed. shCtrl and oeCtrl: negative control lentiviruses. shJMJD3: THP-1 cells infected with lentiviruses to knockdown JMJD3. oeJMJD3: THP-1 cells infected with lentiviruses to overexpress JMJD3. Following lentivirus infection, THP-1 cells were treated with PMA for 24 hours, followed by IL-4 and IL-13 for 48 hours. After this, the medium was replaced by RPMI 1640 medium without FBS for an additional 24 hours. Protein and mRNA were then extracted, and the supernatant was collected. (B) Relative expression of Arg1 mRNA. (C) Concentrations of TNF in the supernatant of different groups of macrophages. (D) Relative expression of IRF4 mRNA. (E, F) Western blot analysis of CD206, JMJD3, and IRF4 protein. The experiments were replicated three times. * p < 0.05, ** p < 0.01, ***p < 0.001, ****p < 0.0001. Abbreviations: JMJD3, Jumonji domain-containing protein 3; Arg1, arginase 1; TNF, tumor necrosis factor; IRF4, interferon regulatory factor 4; CD206, cluster of differentiation 206.
Article Snippet: Subsequently, the cells were treated with 20 ng/ml interleukin-4 (IL-4) (200-04-02; PeproTech, Cranbury, NJ, USA) and 20 ng/ml interleukin-13 (IL-13) (HY- P70568 ; MCE, Monmouth Junction, NJ, USA).
Techniques: Infection, Expressing, Negative Control, Knockdown, Western Blot