Journal: iScience

Article Title: Platelet DKK1 promotes tolerogenic dendritic cells and non-healing responses in cutaneous leishmaniasis

doi: 10.1016/j.isci.2026.115090

Figure Lengend Snippet: Decreased MPO + , CD11b + , and MHC class II + neutrophils in MyD88 (PKO) - and DKK1 (PKO) -infected mice (A–G) Six-week-old female WT-BALB/c, MyD88 (PKO) and DKK1 (PKO) mice were challenged with infective metacyclic promastigote (2 ×10 6 parasites, n = 5 per group) of L . major via the footpad. Non-infected BALB/c mice ( n = 10/2 feet per mouse) were given 0.9% NaCl saline. Cells were isolated from the footpads of all infected and non-infected mice at days 3 and 14 PI. Samples were analyzed by flow cytometry for MPO + , CD11b + , and MHC class II + neutrophils. A representative flow cytometry dot plot of MPO + , CD11b + , and MHC class II + neutrophils on day 3 is presented in . The percentage of MPO + , CD11b + , and MHC class II + cells in the different experimental groups is shown in column graphs (A, B, C, D, E, and F), while the absolute number of neutrophils obtained on day 3 PI is indicated (G). In all experiments, infected and non-infected BALB/c mice served as positive and negative controls, respectively. Results are presented as mean (±SEM) and are representative of two independent experiments. One-way ANOVA followed by Bonferroni’s post hoc test was used to analyze the data ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ns, non-significant ( p > 0.05).

Article Snippet: Rested immature dendritic cells were stimulated with 100 ng/mL of recombinant DKK1 (R & D Systems), 20 ng/mL of recombinant IL-10 (Thermo Fisher Scientific), or 10 ng/mL of recombinant TNF-α (Thermo Fisher Scientific).

Techniques: Infection, Saline, Isolation, Flow Cytometry

Journal: iScience

Article Title: Platelet DKK1 promotes tolerogenic dendritic cells and non-healing responses in cutaneous leishmaniasis

doi: 10.1016/j.isci.2026.115090

Figure Lengend Snippet: Increased CD38 + macrophages and CD8α + dendritic cells in MyD88 (PKO) - and DKK1 (PKO) -infected mice (A–D) Six-week-old female WT-BALB/c, MyD88 (PKO) and DKK1 (PKO) mice were challenged with infective metacyclic promastigote (2 ×10 6 parasites, n = 5 per group) of L . major via the footpad. One week and two weeks post-infection, cells from the infected foot of each mouse in BALB/c, MyD88 (PKO) , DKK1 (PKO) , and non-infected BALB/c mice ( n = 10/2 feet per mouse) were harvested and counted for macrophages (A and B) and dendritic cells (C and D) by flow cytometry. (E–H) Two weeks post-infection, the infected foot from each mouse in BALB/c, MyD88 (PKO) , DKK1 (PKO) ( n = 5 per group), and non-infected BALB/c mice ( n = 10/2 feet per mouse) were examined for CD206 + and CD38 + macrophages (E and F), as well as CD11b + and CD8α + dendritic cells (G and H) by flow cytometry. Representative flow cytometry dot plots showing the analysis of CD206 + , CD38 + macrophages and CD11b + , CD8α + dendritic cells and a dot plot of each sample in all the experimental groups performed on day 14 PI are presented in . Results are presented as mean ± SEM and are representative of two independent experiments. Data analysis was done using one-way ANOVA followed by Bonferroni’s post hoc test ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and “ns” indicates non-significant.

Article Snippet: Rested immature dendritic cells were stimulated with 100 ng/mL of recombinant DKK1 (R & D Systems), 20 ng/mL of recombinant IL-10 (Thermo Fisher Scientific), or 10 ng/mL of recombinant TNF-α (Thermo Fisher Scientific).

Techniques: Infection, Flow Cytometry

Journal: iScience

Article Title: Platelet DKK1 promotes tolerogenic dendritic cells and non-healing responses in cutaneous leishmaniasis

doi: 10.1016/j.isci.2026.115090

Figure Lengend Snippet: DKK1 promotes IL-10 induction in BALB/c-infected mice (A–F) Six-week-old female BALB/c, MyD88 (PKO) and DKK1 (PKO) mice were challenged with infective metacyclic promastigote (2 ×10 6 parasites, n = 5 per group) of L . major via the footpad. Non-infected BALB/c mice ( n = 5) were given 0.9% NaCl saline. Two or fifteen weeks post-infection, cells from draining lymph nodes were isolated. Lymph node cells were incubated with SLAG (50 μg/mL derived from WT parasites). Cell culture supernatant samples obtained were analyzed by ELISA for cytokine production, as shown in the column graphs (A–F). In all the experiments, BALB/c infected and non-infected mice served as positive and negative controls, respectively. Results are presented as mean ± SEM and are representative of two independent experiments. One-way ANOVA followed by Bonferroni’s post hoc test was used to analyze the data ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, “ns” indicates not significant ( p > 0.05).

Article Snippet: Rested immature dendritic cells were stimulated with 100 ng/mL of recombinant DKK1 (R & D Systems), 20 ng/mL of recombinant IL-10 (Thermo Fisher Scientific), or 10 ng/mL of recombinant TNF-α (Thermo Fisher Scientific).

Techniques: Infection, Saline, Isolation, Incubation, Derivative Assay, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Platelet DKK1 promotes tolerogenic dendritic cells and non-healing responses in cutaneous leishmaniasis

doi: 10.1016/j.isci.2026.115090

Figure Lengend Snippet: Impaired CD8 + IL-10 + IFN-γ − , CD4 + IL-10 + IFN-γ − , and CD4 + IL-10 + IFNg + T cells in MyD88 (PKO) - and DKK1 (PKO) -infected mice on day 14 PI (A–M) Six-week-old female BALB/c, MyD88 (PKO) , and DKK1 (PKO) mice were challenged with infective metacyclic promastigote (2 ×10 6 parasites, n = 5 per group) of L . major via the footpad. Non-infected BALB/c mice ( n = 5) were given 0.9% NaCl saline. Two weeks post-infection, the draining and non-draining lymph node cells were isolated. Lymph node cells were incubated with a cell stimulation cocktail for 5 h. After a further 3 h in BFA, cells were stained for intracellular IL-10 and IFN-γ. The stained lymph node cells from each mouse in BALB/c, MyD88 (PKO) - and DKK1 (PKO) -infected mice were determined for the percentage and frequency of CD4 + and CD8 + T cells (A, B, C, and B), percentage and MFI of CD8 + IFN-γ + or CD8 + IL-10 + T cells (E, F, G, and H), percentage and MFI of CD4 + IFN-γ + or CD4 + IL-10 + T cells (I, J, K, and L), and percentage of CD4 + IFN-γ + IL-10 + T cells (M) by flow cytometry. Representative flow cytometry dot plots showing the analyses CD4 + and CD8 + T cells, CD8 + IFN-γ + or CD8 + IL-10 + T cells, CD4 + IFN-γ + or CD4 + IL-10 + T cells, and CD4 + IFN-γ + IL-10 + T cells performed on day 14 PI is indicated ( A). Results are presented as mean ± SEM. and are representative of triplicate experiments. Data analysis was done using one-way ANOVA followed by Bonferroni’s post hoc test ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and “ns” indicates non-significant.

Article Snippet: Rested immature dendritic cells were stimulated with 100 ng/mL of recombinant DKK1 (R & D Systems), 20 ng/mL of recombinant IL-10 (Thermo Fisher Scientific), or 10 ng/mL of recombinant TNF-α (Thermo Fisher Scientific).

Techniques: Infection, Saline, Isolation, Incubation, Cell Stimulation, Staining, Flow Cytometry

Journal: iScience

Article Title: Platelet DKK1 promotes tolerogenic dendritic cells and non-healing responses in cutaneous leishmaniasis

doi: 10.1016/j.isci.2026.115090

Figure Lengend Snippet: Minimal expression and percentage of MHC II + , CD86 + , and CD80 + dendritic cells in the presence of recombinant DKK1 (A) Monocyte-derived dendritic cells were incubated in rDKK1(100 ng/mL), rIL-10 (20 ng/mL) and rTNF-α (10 ng/mL). Cells harvested at 24 and 48 h post-incubation were used to determine the expression and percentage of MHC II + , CD86 + , and CD80 + cells by flow cytometry. Representative flow cytometry dot plots generated 24 h post-incubation showed the analysis of MHC II + , CD86 + and CD80 + as indicated (A). (B–G) The percentage (B, D, and F) and MFI (C, E, and G) of MHC II + , CD86 + and CD80 + in the different experimental conditions are shown in the bar graphs. Results are presented as mean (±SEM) and are representative of triplicate experiments. One-way ANOVA followed by Bonferroni’s post hoc test was used to analyze the data (∗ p < 0.05, ∗∗ p < 0.01; ∗∗∗ p < 0.001; and “ns” indicates non-significant.

Article Snippet: Rested immature dendritic cells were stimulated with 100 ng/mL of recombinant DKK1 (R & D Systems), 20 ng/mL of recombinant IL-10 (Thermo Fisher Scientific), or 10 ng/mL of recombinant TNF-α (Thermo Fisher Scientific).

Techniques: Expressing, Recombinant, Derivative Assay, Incubation, Flow Cytometry, Generated

Journal: iScience

Article Title: Platelet DKK1 promotes tolerogenic dendritic cells and non-healing responses in cutaneous leishmaniasis

doi: 10.1016/j.isci.2026.115090

Figure Lengend Snippet: r-DKK1 blocked TNF-α-induced MHC II + and CD86 + expression of dendritic cells (A) Monocyte-derived dendritic cells were incubated in rDKK1(100 ng/mL), rTNF-α (10 ng/mL), and (rDKK1 + rTNF-α). Cells harvested at 48 h post-incubation were used to determine the percentage of MHC II + and CD86 + cells by flow cytometry. Representative flow cytometry dot plots generated 48 h post-incubation showed the analysis of MHC II + and CD86 + as indicated (A). (B and C) The percentage (B and C) of MHC II + and CD86 + in the different experimental conditions are shown in the bar graphs. Results are presented as mean (±SEM) and are representative of triplicate experiments. One-way ANOVA followed by Bonferroni’s post hoc test was used to analyze the data (∗ p < 0.05, ∗∗ p < 0.01; and “ns” indicates non-significant.

Article Snippet: Rested immature dendritic cells were stimulated with 100 ng/mL of recombinant DKK1 (R & D Systems), 20 ng/mL of recombinant IL-10 (Thermo Fisher Scientific), or 10 ng/mL of recombinant TNF-α (Thermo Fisher Scientific).

Techniques: Expressing, Derivative Assay, Incubation, Flow Cytometry, Generated

Journal: iScience

Article Title: Platelet DKK1 promotes tolerogenic dendritic cells and non-healing responses in cutaneous leishmaniasis

doi: 10.1016/j.isci.2026.115090

Figure Lengend Snippet: Elevated IL-10 and reduced IL-12 production in rDKK1-treated dendritic cells (A and B) Monocyte-derived dendritic cells were incubated in r-DKK1(100 ng/mL), r-IL-10 (20 ng/mL), r-TNF-α (10 ng/mL) or r-TNF-α + r-DKK1. Cell culture supernatants harvested 24 and 48 h post-incubation were used to determine IL-10 and IL-12 production by ELISA, as shown in the bar graphs (A and B). Results are presented as mean ± SEM and are representative of triplicate experiments. One-way ANOVA followed by Bonferroni’s post hoc test was used to analyze the data (∗ p < 0.05, ∗∗ p < 0.01). “ns” indicates not significant.

Article Snippet: Rested immature dendritic cells were stimulated with 100 ng/mL of recombinant DKK1 (R & D Systems), 20 ng/mL of recombinant IL-10 (Thermo Fisher Scientific), or 10 ng/mL of recombinant TNF-α (Thermo Fisher Scientific).

Techniques: Derivative Assay, Incubation, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Platelet DKK1 promotes tolerogenic dendritic cells and non-healing responses in cutaneous leishmaniasis

doi: 10.1016/j.isci.2026.115090

Figure Lengend Snippet: rDKK1-treated dendritic cells significantly increased the percentage of IL-10-producing CD4 T cells (A and B) Rested immature dendritic cells were stimulated with 100 ng/mL of recombinant DKK1 or 10 ng/mL of recombinant TNF-α for 24 h. The rDKK1 and r-TNF-α-treated dendritic cells were incubated for 4 h in a 24-well plate (2 × 10 5 per well) in 200 μL of medium in the presence of SLAG (100 ng/ml). SLAG-loaded rDKK1 and r-TNF-α-treated dendritic cells (2 ×10 5 ) were then washed and co-cultured with infected WT-BALB/c lymph node T cells (1 ×10 6 ) in complete RPMI 1640 medium in a 1:5 ratio for 72 h. The cells were surface-stained with appropriate antibodies before intracellular IL-10 staining with a cell stimulation cocktail and BD Golgi plug. The stained lymph node cells from the experimental condition were determined for the percentage of CD4 + IL-10 + T cells by flow cytometry. Representative flow cytometry dot plots showed the analysis of CD4 + IL-10 + T cells (A). The percentage of CD4 + IL-10 + T cells in the different experimental conditions is indicated (B). The non-treated and r-TNF-α-treated dendritic cells serve as controls. Results are presented as mean ± SEM and are representative of three replicate experiments. One-way ANOVA followed by Bonferroni’s post hoc test was used to analyze the data ∗∗∗ p < 0.001; ns indicates not significant ( p > 0.05).

Article Snippet: Rested immature dendritic cells were stimulated with 100 ng/mL of recombinant DKK1 (R & D Systems), 20 ng/mL of recombinant IL-10 (Thermo Fisher Scientific), or 10 ng/mL of recombinant TNF-α (Thermo Fisher Scientific).

Techniques: Recombinant, Incubation, Cell Culture, Infection, Staining, Cell Stimulation, Flow Cytometry

Journal: iScience

Article Title: Platelet DKK1 promotes tolerogenic dendritic cells and non-healing responses in cutaneous leishmaniasis

doi: 10.1016/j.isci.2026.115090

Figure Lengend Snippet: Lesion size and parasite burden decreased in MyD88 (PKO) - and DKK1 (PKO) -infected mice (A–C) Six-week-old female WT-BALB/c, MyD88 (PKO) and DKK1 (PKO) mice were challenged with infective metacyclic promastigote (2 ×10 6 parasites) of L . major via the footpad. The infected foot from each mouse in BALB/c, MyD88 (PKO) - and DKK1 (PKO) -infected mice ( n = 5 per group) were measured for lesion size weekly using a vernier caliper (A), and parasite burden (at day week 6 and 15PI) was determined by limiting dilution assay (B and C). Results are presented as mean ± SEM. For Figure (A), mice in each infected group were compared with the non-infected group and data analysis was done using one-way ANOVA followed by Bonferroni’s post hoc test ∗ p < 0.05; ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Rested immature dendritic cells were stimulated with 100 ng/mL of recombinant DKK1 (R & D Systems), 20 ng/mL of recombinant IL-10 (Thermo Fisher Scientific), or 10 ng/mL of recombinant TNF-α (Thermo Fisher Scientific).

Techniques: Infection, Limiting Dilution Assay

Journal: Scientific Reports

Article Title: Wnt-3a exacerbates production of TNF-α in LPS stimulated microglia independent of the β-catenin canonical pathway

doi: 10.1038/s41598-026-37653-1

Figure Lengend Snippet: Canonical pathway response to Wnt-3a stimulation. Relative changes and corresponding electropherograms and Wes-generated pseudobands of ( A ) total β-catenin ( B ) active β-catenin in microglia stimulated with LPS and/or Wnt-3a ( n = 5–7/group). ( C ) Analysis of TNF-α levels in microglia stimulated with DKK1 or PBS vehicle via ELISA ( n = 5/group). All values are shown as mean ± standard error of the mean (SEM). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Mouse recombinant DKK1 (20 μg/mL, R&D Systems) was added to inhibit the canonical pathway, while the JNK inhibitor SP600125 (15 μg/mL, Sigma Aldrich) was added to inhibit the PCP pathway.

Techniques: Generated, Enzyme-linked Immunosorbent Assay

Journal: Current Issues in Molecular Biology

Article Title: Microcirculation-Promoting Effect of Escin on Cutaneous Tissue via Gsk3β Down-Regulation

doi: 10.3390/cimb47100840

Figure Lengend Snippet: Escin-activated Wnt/β-Catenin signaling via Gsk3β. BMP2 gene expression, a gene activated by both ( b , d , f ) Wnt3a and ( a , c , e ) escin, ws examined with co-treatment of Wnt signaling inhibitors; ( a , b ) DKK1, ( c , d ) IWR1, and ( e , f ) Wnt-C59. Control, non-treated control; significantly different compared with each Wnt3a or Escin treated group without inhibitors (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Human recombinant DKK1 were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Gene Expression, Control