Fig. 1 | Communications Biology

Fig. 1

From: Wnt5a is a TLR2/4-ligand that induces tolerance in human myeloid cells

Fig. 1

Wnt5a expression is induced by endotoxin and promotes anti-inflammatory IL-10 in primary human myeloid cells. a Wnt5a expression in primary human monocytes (left, mRNA as assessed by RT-qPCR) and primary human monocyte derived M1 macrophages (right, protein as assessed by western blotting). Wnt5a or LPS (24 h) in the absence of serum (serum free; SF). (n = 3) Dunnett’s test was used for multiple comparisons. b LPS induces Wnt5a expression in human primary human alveolar macrophages, as evidenced by analysis of published GEO datasets. Mann–Whitney t-test. c Wnt5a induces anti-inflammatory cytokines in primary human monocytes. Primary human monocytes from healthy controls were treated with rWnt5a and/or LPS (6 h SF). The levels of secreted IL-10, TNFα, and IL-6 were analysed by CBA (n = 5); IL-8 by ELISA (n = 3). ANOVA (no brackets) or Student’s t-test (brackets). For multiple comparisons Dunn’s test was used. d Wnt5a induces transcription of IL10 mRNA in primary human monocytes. (n = 3) Dunnett’s test was used for multiple comparisons. e Wnt5a does not enhance IL10 mRNA stability in primary human monocytes. Actinomycin D (ActD). The mRNA levels were determined using RT-qPCR. (n = 9) Dunnett’s test was used for multiple comparisons. f Wnt5a induces activation of the IL10 promoter. Luciferase IL-10 promoter assay was performed using human THP-1 cells transfected with HA-Wnt5a or stimulated with LPS (18 h). Relative dual luciferase units (RLU) were determined, with the pRL-TK vector as the control. (n = 7) Dunnett’s test was used for multiple comparisons. g Overexpression of pcDNA3-HA-Wnt5a (48 h) in THP-1 cells induces cytokine expression. Cytokines were analysed by CBA. (n = 7) Dunnett’s test was used for multiple comparisons. h Wnt5a does not induce NFκB activity. THP-1- Blue™ NFκB cells were treated with rWnt5a and/or LPS (6 h). (n = 6). Dunnett’s test was used for multiple comparisons. i Overexpression of pcDNA3-HA-Wnt5a (48 h) in THP1-DualTM MyD-KO cells reduces cytokine expression as compared to g. Cytokines were analysed by CBA. (n = 4) Dunnett’s test was used for multiple comparisons. Control cells were transfected with an anti-sense pcDNA3-as-Wnt5a vector. Error bars indicate SEM; *p < 0.05, **p < 0.01, and ***p < 0.001. Data were analysed by ANOVA or t-test as indicated

Back to article page