Journal: bioRxiv

Article Title: Improved efficacy and tolerability of antisense oligonucleotide with Guanidine- bridged nucleic acid

doi: 10.64898/2026.01.08.698310

Figure Lengend Snippet: GuNA suppressed immunotoxicity. (A) Amount of IL-6 release relative to untreated control. Cells were treated with modified gapmer by gymnotic uptake for 24 hours. (n=3) (B) Percentage of CCL22 mRNA expression relative to untreated control. Cells were treated with modified gapmer by gymnotic uptake for 24 hours. (n=2)

Article Snippet: The TaqMan probe product IDs were human ACTB (Hs99999903_m1), human APOB (Hs00181142_m1), human SOD1 (Hs00533490_m1), human CCL22 (Hs01574247_m1), mouse Gapdh (Mm99999915_g1), mouse Malat1 (Mm01227912_s1), and mouse Cd68 (Mm03047343_m1).

Techniques: Control, Modification, Expressing

Journal: Cell Biology and Toxicology

Article Title: SENP3 mediated DeSUMOylation of macrophage derived CCL17 accelerates atherosclerosis via regulation of Treg

doi: 10.1007/s10565-025-10099-3

Figure Lengend Snippet: Macrophages stimulated by oxLDL affect the chemotaxis of Treg through CCL17 and CCL22 competition. Macrophages were stimulated with oxLDL, and Treg cells were co-cultured with macrophage supernatant. A Immunofluorescence detection of co-localization of CCL22 and macrophages (CD68) in plaque tissue of AS mice. B The content of CCL22 in the supernatant of macrophages was detected by ELISA. C Transwell detected the effect of macrophages on the chemotaxis of Treg. ( D - E ) Transwell detected the migration of Treg cells. * p < 0.05, ** p < 0.01, *** p < 0.001. N = 3

Article Snippet: Prior to the transwell migration assay, the induced Tregs were treated with recombinant mouse CCL17 (MCE, HY-P71891A) or recombinant mouse CCL22 (MCE, HY-P7248) or anti-CCL17 (R&D, Catalog #: MAB529) and anti-CCL22 antibodies (R&D, Catalog #: MAB529).

Techniques: Chemotaxis Assay, Cell Culture, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Migration

Journal: The EMBO Journal

Article Title: DNGR-1 signalling limits dendritic cell activation for optimal antigen cross-presentation

doi: 10.1038/s44318-025-00620-z

Figure Lengend Snippet: ( A ) Aligned DNGR-1 (CLEC9A, C9; left) or Dectin-1 (CLEC7A, C7; right) cytoplasmic domain sequences from indicated species. HemITAM sequence is bolded in red. Consensus is represented by an asterisk (*) and similarity with a dot (.). ( B ) Aligned hemITAM motifs of DNGR-1 and Dectin-1 from Mus musculus . The triacidic motif from Dectin-1 is also depicted with the corresponding sequence from DNGR-1. ( C ) Schematic of chimeric receptor comprised of the intracellular cytoplasmic domain (ICD) of Dectin-1 fused with the transmembrane (TM) and extracellular domain (ECD) of DNGR-1 (C7::C9), or I6G DNGR-1 (C9(I6G)) transduced into C9 KO MuTuDC (left). Right side depicts WT Dectin-1 (C7), or chimeric receptor constructs using the TM and ECD of Dectin-1 fused with the ICD of WT (C9::C7), I6G (C9(I6G)::C7), or Y7F (C9(Y7F)::C7) DNGR-1 transduced into RAW 264.7 cells. ( D , E ) Representative flow cytometric analysis of surface proteins from C9 KO MuTuDCs or those reconstituted with indicated receptors and stimulated overnight ± with DNGR-1L. ( E ) Quantification of flow cytometric analysis of surface proteins from MuTuDCs in ( D ), as well as CCL22 in cultured supernatants from those cells. Mean ± SD from biological duplicates is plotted. ( F ) Representative histograms and quantification of flow cytometric analyses of surface marker MFI or TNF-α in cultured supernatants from RAW 264.7 cells ectopically expressing indicated receptors or transduced with empty vector (EV) stimulated overnight ± with zymosan depleted (Zym-D). Mean ± SD from biological duplicates is plotted. ( G ) RNAseq volcano plot of differentially expressed genes between 12.5 nM DNGR-1L stimulated versus untreated (Control) KO/C9 (left) or KO/C9(I6G) MuTuDCs (right). N = 3 per group. ( H ) RT-qPCR analysis of indicated genes from MuTuDCs treated with 12.5 nM DNGR-1L over time. Mean ± SD from biological duplicates is plotted. ( I ) Gene set enrichment analysis (GSEA) of Reactome signalling pathways identified in DNGR-1L-stimulated KO/C9(I6G) MuTuDCs versus KO/C9 MuTuDCs from ( G ). Data are representative of two ( F , H ), or three ( D , E ) independent experiments. See also Fig. . .

Article Snippet: Taqman Ccl22 , Thermo Fisher Scientific , Assay ID: Mm00436438_m1.

Techniques: Sequencing, Construct, Cell Culture, Marker, Expressing, Transduction, Plasmid Preparation, Control, Quantitative RT-PCR

Journal: The EMBO Journal

Article Title: DNGR-1 signalling limits dendritic cell activation for optimal antigen cross-presentation

doi: 10.1038/s44318-025-00620-z

Figure Lengend Snippet: ( A ) Analysis of the indicated surface marker expression (top, middle; flow cytometry) or CCL22 released into cultured supernatants (bottom; ELISA) from C9 KO MuTuDCs reconstituted or not with the indicated receptors and stimulated overnight ± DNGR-1L. Mean ± SEM from biological replicates pooled from two independent experiments (left) and representative flow cytometry profiles (right) are plotted. Data here are partly represented in Fig. and are relative to average of untreated controls to better emphasise the response to DNGR-1L. Dotted line represents 1. ( B ) Flow cytometric analysis of surface DNGR-1 (C9) expression in C9 KO MuTuDCs or those reconstituted or not with the indicated receptors. Cell lines were established after sorting for equal expression of DNGR-1. ( C ) Flow cytometric analysis of surface Dectin-1 (C7) expression by parental RAW 264.7 cells or cells ectopically expressing the indicated receptors or transduced with empty vector (EV). Cell lines were established after sorting for equal expression of Dectin-1. ( B , C ) Representative histograms (left) and mean MFI ± SEM (right) are plotted from biological ( B ) quintuplets or ( C ) triplicates. ( D ) Analysis of the indicated surface marker expression (top, middle; flow cytometry) or TNF-α released into cultured supernatants (bottom; ELISA) from RAW 264.7 cells ectopically expressing the indicated receptors or transduced with EV and stimulated overnight ± Zym-D. Mean ± SEM from biological replicates pooled from two independent experiments (left) and representative flow cytometry profiles (right) are plotted. Data here are partly represented in Fig. and are relative to average of untreated controls to better emphasise the response to Zym-D. Dotted line represents 1. Data are representative of two ( D ), or three ( A – C ) independent experiments. Data were analysed using Tukey-corrected two-way ANOVA with significant values comparing against untreated samples plotted ( A , D ). ( A ) I-A/E hi CD86 hi * P = 0.0117, **** P < 0.0001; CCR7 * P = 0.0279 (KO/C7::C9), P = 0.0134 (KO/C9(I6G)), **** P < 0.0001; CCL22 **** P < 0.0001, ( D ) H2-D d ** P = 0.0011, **** P < 0.0001; CCR7 * P = 0.0319, ** P = 0.0040, **** P < 0.0001; TNF-α ** P = 0.0033, *** P = 0.0004, **** P < 0.0001.

Article Snippet: Taqman Ccl22 , Thermo Fisher Scientific , Assay ID: Mm00436438_m1.

Techniques: Marker, Expressing, Flow Cytometry, Cell Culture, Enzyme-linked Immunosorbent Assay, Transduction, Plasmid Preparation

Journal: The EMBO Journal

Article Title: DNGR-1 signalling limits dendritic cell activation for optimal antigen cross-presentation

doi: 10.1038/s44318-025-00620-z

Figure Lengend Snippet: ( A , B ) Flow cytometric analyses of surface proteins and ELISA of indicated proteins released in culture supernatants from ( A ) RAW 264.7 cells ectopically expressing indicated receptors or transduced with EV stimulated overnight ± Zym-D or ( B ) C9 KO MuTuDCs reconstituted with indicated receptors stimulated overnight ± DNGR-1L. Data shown as mean ± SEM from biological triplicates. Note that the TNF-α data in ( A ) are plotted in two different ways (absolute versus relative concentration to untreated controls) to emphasise the similarity between C7 and C9(I6G)::C7. ( C ) Uptake of DNGR-1L-coupled beads by C9 KO MuTuDCs or those reconstituted with indicated receptors assessed by flow cytometry. Plotted as phagocytic index (% internalised beads x bead MFI of bead + cells/arbitrary unit) mean ± SD from biological duplicates. ( D , E ) C9 KO MuTuDCs reconstituted with C7::C9 or C7(G14I)::C9 receptors co-cultured with DNGR-1L-OVA coupled beads. Single internalised bead + cells were subsequently sorted and co-cultured with OT-I T E cells. ( D ) Schematic of experiment (left) and representative flow plots from pre- and post-sort enrichment (right). ( E ) ELISA for IFN-γ released from OT-I T E cells co-cultured with MuTuDCs from ( D ) or MuTuDCs loaded with exogenous SIINFEKL peptide. Mean ± SD from biological duplicates is plotted. Data are representative of two ( D , E ) or ≥ three ( A – C ) independent experiments. Data were analysed using Tukey-corrected two-way ANOVA ( A – C , E ). Significant values comparing against untreated controls ( A , B ) or between KO/C7(G14I)::C9 ( E ) are plotted. ( A ) H2-D d ** P = 0.0043, *** P = 0.0006, **** P < 0.0001; CD83 * P = 0.0374, *** P = 0.0002, **** P < 0.0001; TNF-α ** P = 0.0017 (C7), P = 0.0067 (C9(Y7F)::C7, *** P = 0.0004 (C7(G14I)), P = 0.0006 (C9::C7), **** P < 0.0001, ( B ) CD86 * P = 0.0469, **** P < 0.0001; CCR7 ** P = 0.0097, *** P = 0.0004, **** P < 0.0001; CCL17 **** P < 0.0001; CCL22 * P = 0.0217, ** P = 0.0018, **** P < 0.0001, ( C , E ) **** P < 0.0001. See also Fig. . .

Article Snippet: Taqman Ccl22 , Thermo Fisher Scientific , Assay ID: Mm00436438_m1.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Transduction, Concentration Assay, Flow Cytometry, Cell Culture