Journal: Oncotarget

Article Title: C1q/Tumor necrosis factor-related protein-3 protects macrophages against LPS-induced lipid accumulation, inflammation and phenotype transition via PPARγ and TLR4-mediated pathways

doi: 10.18632/oncotarget.19657

Figure Lengend Snippet: (A) Impacts of increasing CTRP3 concentration on the viability of THP-1 differentiated macrophages and mouse peritoneal macrophages. Cells were suspended into 1 × 10 4 cells /100μL/well; THP-1 cells were induced to differentiate into macrophages by culture with 100 ng/mL PMA for 48 h. Mouse peritoneal macrophages were extracted and cultured as mentioned above, after which increasing doses of CTRP3 (0-10 μg/mL) were added; 24 h later, the CCK-8 assay was used to detect viability (mean ± SD). (B-C) Impacts of CTRP3 on LPS-triggered macrophage lipid deposition and foam cell formation in both kinds of macrophages (1 × 10 6 cells/mL/well). The cholesterol content was detected using a cholesterol quantification kit to evaluate lipid deposition in LPS-triggered macrophages (B, mean ± SD, * : compared to the control group, P < 0.05; # : compared to the ox-LDL and LPS treatment group, P < 0.05). Oil red O staining was performed to analyze the effect of CTRP3 on foam cell formation (C, THP-1 cells: 200×, mouse peritoneal macrophages: 400×). At least three independent experiments were performed.

Article Snippet: Human recombinant CTRP3 protein obtained using an in vitro wheat germ expression system was purchased from Abnova (Taipei, Taiwan).

Techniques: Concentration Assay, Cell Culture, CCK-8 Assay, Control, Staining

Journal: Oncotarget

Article Title: C1q/Tumor necrosis factor-related protein-3 protects macrophages against LPS-induced lipid accumulation, inflammation and phenotype transition via PPARγ and TLR4-mediated pathways

doi: 10.18632/oncotarget.19657

Figure Lengend Snippet: THP-1 cells and mouse peritoneal macrophages (1 × 10 6 cells/mL/well) were preincubated with different concentrations of CTRP3 (0-1.0 μg/ mL) for 30 min. After LPS and ox-LDL stimulation, the supernatant protein levels of TNFα, IL-6, MCP-1, IL-1β and MMP-9 were evaluated by ELISA. At least three independent experiments were performed (mean ± SD, * : compared to the control group, P < 0.05, # : compared to the LPS and ox-LDL treatment group, P < 0.05).

Article Snippet: Human recombinant CTRP3 protein obtained using an in vitro wheat germ expression system was purchased from Abnova (Taipei, Taiwan).

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Oncotarget

Article Title: C1q/Tumor necrosis factor-related protein-3 protects macrophages against LPS-induced lipid accumulation, inflammation and phenotype transition via PPARγ and TLR4-mediated pathways

doi: 10.18632/oncotarget.19657

Figure Lengend Snippet: Cells (1 × 10 6 cells /mL/well) were treated, and expression of specific M1 and M2 markers on the cell surface was detected by FCM to observe macrophages polarization. For THP-1 cells, cells expressing CD68 but not CD206/MR (CD68 + CD206 − ) were considered to be M1 macrophages; cells co-expressing CD68 and CD206 (CD68 + CD206 + ) were considered to be M2 macrophages. For mouse peritoneal macrophages, cells expressing CCR7 and CD86 were considered to be M1 macrophages; cells expressing CD163 and CD206/MR were considered to be M2 macrophages. The macrophage phenotype ratio was calculated as (CCR7 plus CD86 expression)/(CD163 plus CD206 expression). At least three independent experiments were performed (mean ± SD, * : compared to the control group, P < 0.05, # : compared to the LPS but without CTRP3 treatment group, P < 0.05).

Article Snippet: Human recombinant CTRP3 protein obtained using an in vitro wheat germ expression system was purchased from Abnova (Taipei, Taiwan).

Techniques: Expressing, Control

Journal: Oncotarget

Article Title: C1q/Tumor necrosis factor-related protein-3 protects macrophages against LPS-induced lipid accumulation, inflammation and phenotype transition via PPARγ and TLR4-mediated pathways

doi: 10.18632/oncotarget.19657

Figure Lengend Snippet: PMA-induced THP-1 macrophages (1 × 10 6 cells/mL/well) were divided into three groups: the control group was treated with PBS for 24 h; the LPS group was treated with 100 ng/mL LPS for 24 h; the LPS+CTRP3 group was treated with 1 μg/mL CTRP3 for 30 min before LPS stimulation for 24 h. mRNA expression of specific M1 marker and M2 markers was assessed by qRT-PCR. Relative mRNA concentrations were calculated using the ΔΔCT method, and the expression level was normalized to that of endogenous β-actin. At least three independent experiments were performed (mean ± SD, * : compared to the control group, P < 0.05, # : compared to the LPS group, P < 0.05).

Article Snippet: Human recombinant CTRP3 protein obtained using an in vitro wheat germ expression system was purchased from Abnova (Taipei, Taiwan).

Techniques: Control, Expressing, Marker, Quantitative RT-PCR

Journal: Oncotarget

Article Title: C1q/Tumor necrosis factor-related protein-3 protects macrophages against LPS-induced lipid accumulation, inflammation and phenotype transition via PPARγ and TLR4-mediated pathways

doi: 10.18632/oncotarget.19657

Figure Lengend Snippet: Different concentrations of CTRP3 were added before cells (1 × 10 6 cells/mL/well) were exposed to LPS and ox-LDL, and protein expression of TLR4, MyD88, p-NF-κB p65 and NF-κB p65 was assessed by WB (A-D) . Protein expression of PPARγ, LXRα, ABCA1 and ABCG1 was assessed by WB (E-I) (mean ± SD, * : compared to the control group, P < 0.05, # : compared to the LPS and ox-LDL treatment group, P < 0.05).

Article Snippet: Human recombinant CTRP3 protein obtained using an in vitro wheat germ expression system was purchased from Abnova (Taipei, Taiwan).

Techniques: Expressing, Control

Journal: Oncotarget

Article Title: C1q/Tumor necrosis factor-related protein-3 protects macrophages against LPS-induced lipid accumulation, inflammation and phenotype transition via PPARγ and TLR4-mediated pathways

doi: 10.18632/oncotarget.19657

Figure Lengend Snippet: PMA-induced THP-1 macrophages (1 × 10 6 cells/mL/well) were divided into five groups:the control group was treated with PBS; the LPS group was treated with 100 ng/mL LPS for 24 h prior to 100 μg/mL ox-LDL for 24 h; the LPS+CTRP3 (CT3) group was pretreated with 1 μg/mL CTRP3 for 30 min before LPS and ox-LDL stimulation; the LPS+PDTC group was pretreated with 100 μM PDTC for 1 h and then stimulated with LPS and ox-LDL; the LPS+PDTC+CT3 group was treated with 100 μM PDTC for 1 h and then 1 μg/mL CTRP3 for 30 min before LPS and ox-LDL stimulation. Expression of TLR4, MyD88, NF-κB p-p65, t-p65 and β-actin was detected by WB (mean ± SD, * : compared to the control group, P < 0.05, # : compared to the LPS group, P < 0.05, $ : compared to the LPS+CT3 group, P < 0.05).

Article Snippet: Human recombinant CTRP3 protein obtained using an in vitro wheat germ expression system was purchased from Abnova (Taipei, Taiwan).

Techniques: Control, Expressing

Journal: Oncotarget

Article Title: C1q/Tumor necrosis factor-related protein-3 protects macrophages against LPS-induced lipid accumulation, inflammation and phenotype transition via PPARγ and TLR4-mediated pathways

doi: 10.18632/oncotarget.19657

Figure Lengend Snippet: PMA-induced THP-1 macrophages (1 × 10 6 cells/mL/well) were divided into six groups: the first three group were treated as mentioned above, and the other three groups were transfected with si-ctrl, si-PPARγ or si-LXRα for 48 h, and then treated with 1 μg/mL CTRP3 for 30 min, and finally stimulated with LPS and ox-LDL. Expression of PPARγ, LXRα, ABCA1, ABCG1 and β-actin was detected by WB (mean ± SD, * : compared to the control group, P < 0.05, # : compared to the LPS group, P < 0.05, $ : compared to the LPS+CT3 group, P < 0.05).

Article Snippet: Human recombinant CTRP3 protein obtained using an in vitro wheat germ expression system was purchased from Abnova (Taipei, Taiwan).

Techniques: Transfection, Expressing, Control