Journal: bioRxiv

Article Title: Mapping the MIF-2 Chemokine Interactome Reveals MIF-2–CCL20 Complex Formation in Liver Fibrosis

doi: 10.64898/2026.03.02.708954

Figure Lengend Snippet: Shown are gene expression data re-analyzed from the GSE135251 dataset, focusing on CCL20, MIF, and MIF-2 (DDT) together with their corresponding receptors CCR6, CXCR4, and CD74 in NASH across increasing fibrosis stages (F0–F4; shades of magenta) compared with NAFL (grey) ( Govaere et al ., 2020 ). Statistical comparisons between groups were performed using the Mann–Whitney U test, with P values adjusted for multiple testing using the Holm–Bonferroni correction. NASH: non-alcoholic steatohepatitis; NAFL: non-alcoholic fatty liver disease. Statistical significance is indicated by actual adjusted P values.

Article Snippet: Antibodies included APC-conjugated anti-CXCR4 (BioLegend; Cat# 306510), FITC-conjugated anti-CD74 (BD Biosciences; Cat# 555540), and anti-CCR6 antibodies (PE-conjugated, Miltenyi; Cat# 130-100-377; APC/Cy7-conjugated, BioLegend; Cat# 353432).

Techniques: Gene Expression, MANN-WHITNEY

Journal: Biochemistry and Biophysics Reports

Article Title: Comparative analysis of activation of macrophages/microglia in diabetic retinopathy and Familial Exudative Vitreoretinopathy

doi: 10.1016/j.bbrep.2025.102396

Figure Lengend Snippet: FZD4 knockout mice develop FEVR features. FZD4 knockout mice (FZD4KO) was used as an FEVR model. (A) Western blot for FZD4 in the retinal tissue of FZD4 knockout mice, compared to wildtype controls (WT). (B–C) Electroretinogram (ERG) assessments at 16 weeks, shown by representative chars (B) and by quantification (C). Blue circle: b-wave; green circle: a-wave. (D) Retinal vascular density by CD31 + area quantification. N = 5. ∗p < 0.05. NS: no significant.

Article Snippet: Knockout of FZD4 in these mice were validated by Western blot for FZD4 in the retinal tissue, using monoclonal antibody against mouse FZD4 (MAB195–050, R&D Systems).

Techniques: Knock-Out, Western Blot

Journal: Life Science Alliance

Article Title: TRPM7 and magnesium orchestrate human CD4 T-cell activation and differentiation

doi: 10.26508/lsa.202503357

Figure Lengend Snippet: (A) Schematic description of naïve CD4 T-cell differentiation towards FOXP3-expressing regulatory T cells and RORƔt-expressing T H 17 cells, including respective cytokine polarization milieus. (B, C) Percentages of CD45RA − cells and (C) CD25 + CD127 lo cells upon polarization of naïve CD4 T cells toward iT reg cells in various NS8593 concentrations (red) compared with DMSO control (Ctrl, black), n = 6–7. (D, E) Representative FACS histograms and (E) quantification of FOXP3 expression levels of CD25 + CD127 lo iT reg cells upon 6 d polarization of naïve CD4 T cells in presence of various NS8593 concentrations (red) or DMSO control (Ctrl, black), n = 6–7. (F, G) Percentages of CD45RA − cells and (G) CD25 + CD127 lo cells upon polarization of naïve CD4 T cells toward iT reg cells in presence of 6 mM MgCl 2 (MgCl 2 , blue) compared with H 2 O control (Ctrl, black), n = 7. (H, I) Representative FACS histograms and (I) quantification of FOXP3 expression levels of CD25 + CD127 lo iT reg cells upon 6 d polarization of naïve CD4 T cells in presence of 6 mM MgCl 2 (MgCl 2 , blue) compared with H 2 O control (Ctrl, black), n = 6. (J) Percentages of CCR6 + cells upon polarization of naïve CD4 T cells towards iT H 17 cells in presence of various NS8593 concentrations (red) compared with DMSO control (Ctrl, black), n = 6. (K, L) Representative FACS histograms and (L) quantification of RORƔt expression levels of CCR6 + iT H 17 cells upon 6 d polarization of naïve CD4 T cells in presence of various NS8593 concentrations (red) or DMSO control (Ctrl, black), n = 4–6. (M) Percentages of CCR6 + cells upon polarization of naïve CD4 T cells towards iT H 17 cells in presence of 6 mM MgCl 2 (MgCl 2 , blue) compared with H 2 O control (Ctrl, black), n = 6. (N, O) Representative FACS histograms and (O) quantification of RORƔt expression levels of CCR6 + iT H 17 cells upon 6 d polarization of naïve CD4 T cells in presence of 6 mM MgCl 2 (MgCl 2 , blue) compared with H 2 O control (Ctrl, black), n = 5. (P) Graphical summary of TRPM7-(in)dependent T-cell activation and differentiation towards iT reg and iT H 17 cells. Pharmacological blockade of TRPM7 reduces intracellular Mg 2+ levels, leads to reduced Ca 2+ signaling and results in reduced IL-2 secretion, impaired up-regulation of T-cell activation markers CD69 and CD25, and diminished proliferation upon TCR stimulus (left). TRPM7 inhibition during polarization of naïve CD4 T cells into iT reg cells preserves FOXP3 + signals of CD25 + CD127 lo iT reg cells. Polarization of naïve CD4 T cells into iT H 17 cells results in augmented RORƔt expression in the presence of 6 mM Mg 2+ , which is reduced upon TRPM7 inhibition, highlighting the need for Mg 2+ uptake and related TRPM7-dependent intracellular signaling for iT H 17 cell polarization (right). (B, C, E, F, G, I, J, L, M, O) Statistics: one-way ANOVA (B, C, E, J, L) and t test (F, G, I, M, O). * P < 0.05; ** P < 0.005; *** P < 0.0005; **** P < 0.0001 and n.s., not significant. Data are mean ± SD.

Article Snippet: Surface staining was performed using the following antibodies: α-human CD25-PE (BC96; BioLegend), α-human CCR6-PEVio770 (REA190; Milteny).

Techniques: Cell Differentiation, Expressing, Control, Activation Assay, Inhibition

Journal: Life Science Alliance

Article Title: TRPM7 and magnesium orchestrate human CD4 T-cell activation and differentiation

doi: 10.26508/lsa.202503357

Figure Lengend Snippet: (A) Representative FACS plots and gating strategy for iT H 17 cells after 6 d of differentiation of naïve CD4 T cells, shown for Ctrl cells. Naïve CD4 T cells were used for gating. (B) Percentage of viable cells upon polarization of naïve CD4 T cells toward T H 17 cells in various NS8593 concentrations (red) compared with DMSO control (Ctrl, black), gated as in (A), n = 6. (C) Percentage of viable cells upon polarization of naïve CD4 T cells towards T H 17 cells in presence of 6 mM MgCl 2 (MgCl 2, blue) compared with H 2 O control (Ctrl, black), gated as in (A), n = 6. (D, E, F) Percentages of viable cells (D), CCR6 + cells (E) and RORƔt expression (F) upon polarization of naïve CD4 T cells toward T H 17 cells in presence of 1 μM Apamin (Apamin, blue) compared with DMSO control (Ctrl, gray), gated as in (A), n = 5–6. (B, C, D, E, F) Statistics: one-way ANOVA (B) and t test (C, D, E, F). **** P < 0.0001 and n.s., not significant. Data are mean ± SD.

Article Snippet: Surface staining was performed using the following antibodies: α-human CD25-PE (BC96; BioLegend), α-human CCR6-PEVio770 (REA190; Milteny).

Techniques: Control, Expressing