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: THP-1 cells were treated under different conditions: PMA (50 ng/ml) alone, or PMA (50 ng/ml) for 24 hours, followed by a medium change and subsequent treatment with IL-4 (20 ng/ml) and IL-13 (20 ng/ml) for 48 hours. After this, the medium was replaced with RPMI 1640 medium without FBS for 24 hours. Protein and mRNA were then extracted, and the supernatant was collected. (A) Morphological features of THP-1 cells under different conditions. Left: untreated THP-1 cells. Middle: PMA (50 ng/ml) for 24 hours. Right: PMA for 24 hours, followed by IL-4 and IL-13 (20 ng/ml) and IL-13 (20 ng/ml) for 48 hours. Scale bar: 50 μm. (B) Relative expression of Arg1 mRNA. (C) Relative expression of Arg1 protein. (D) Concentrations of TNF and IL-10 in the supernatant. (E) Relative mRNA expression of JMJD3 and IRF4. (F) Protein expression of JMJD3, p-STAT6-Y641, and IRF4. The experiments were replicated three times. * p < 0.05, ** p < 0.01, ***p < 0.001, ****p < 0.0001. Abbreviations: Arg1, arginase 1; PMA, Phorbol 12-myristate 13-acetate; IL-10, interleukin-10; TNF, tumor necrosis factor; JMJD3, Jumonji domain-containing protein 3; IRF4, interferon regulatory factor 4; p-STAT6, phosphorylated signal transducer and activator of transcription 6.

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: Expressing

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

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: THP-1 cells were first treated with PMA for 24 hours. Following this, AS1517499 (1μM) was administered for 30 minutes, and then the cells were treated with IL-4 and IL-13. 48 hours later, the medium was replaced with RPMI 1640 medium without FBS and cells were incubated for an additional 24 hours. Protein and mRNA were then extracted, and the supernatant was collected. (A) The mRNA expression of Arg1. (B) Concentrations of TNF in the supernatant of different groups of macrophages. (C) Concentrations of IL-10 in the supernatant of different groups of macrophages. (D) Expression of IRF4 mRNA. (E) Expression of JMJD3 mRNA. (F) Western blot analysis of JMJD3, p-STAT6, and IRF4 protein expression. 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; IL-10, interleukin-10; p-STAT6, phosphorylated signal transducer and activator of transcription 6.

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: Incubation, Expressing, Western Blot