Journal: Blood Vessels, Thrombosis & Hemostasis

Article Title: A second generation of C3 humanized rats for preclinical evaluation of human C3 inhibitors

doi: 10.1016/j.bvth.2026.100138

Figure Lengend Snippet: The second-generation hC3 rats express human, but not rat C3 in their plasma and tissues, at levels higher than those in the first-generation hC3 rats. (A) Plasma from both human and rat samples were collected, and the presence of human (hC3) and rat C3 (rC3) were evaluated by western blots. Purified human C3 (hC3), rat C3 (rC3), human C3 depleted serum (hC3-dpl), and C3 knockout rat serum rat plasma (rC3-KO) were used as controls. Arrows pointed to the human or rat C3 band. (B) Plasma hC3 levels in the second-generation hC3 rats (both male and female) were evaluated using a human C3 ELISA kit (Hycult) and compared to the levels in the first-generation hC3 rat plasma. (C) Total RNA was extracted from the liver, retina, spleen, kidney, and brain of WT, second-generation, and first-generation hC3 rats, and then semiquantitative reverse transcription PCR was used to detect and quantitate levels of human C3 (hC3) transcripts using the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as internal controls. 1st gen, first generation; 2nd gen, second generation.

Article Snippet: Blood levels of human C3 protein were measured in the identified human C3 Tg rats using an enzyme-linked immunosorbent assay (ELISA) kit (Hycult, catalog no. HK366) to identify the founder rats with close-to-physiological expression levels of human C3.

Techniques: Clinical Proteomics, Western Blot, Purification, Knock-Out, Enzyme-linked Immunosorbent Assay, Reverse Transcription

Journal: Bioactive Materials

Article Title: Splenic dendritic cell-targeting mRNA transfection of H-type ionizable lipid-based LNPs for enhancing tumor immunotherapy

doi: 10.1016/j.bioactmat.2026.02.018

Figure Lengend Snippet: In vivo DC maturation and T cell activation in C57BL/6J mice induced by mOVA/H 18 NPs through intravenous injection. C57BL/6J mice were vaccinated with different formulations on Day 0 and Day 5. On Day 10, the spleens of mice were collected and analyzed by flow cytometry. Quantification analysis of (A) CD80 + CD86 + DCs and (B) CD40 + DCs in the spleen. Quantification analysis of (C) CD3 + CD4 + T cells and (D) CD3 + CD8 + T cells in the spleen. (E) Quantification analysis and (F) representative flow cytometry contour plots of OVA-specific CD8 + T cells among all cell populations in the spleen. (G) Quantification analysis and (H) representative flow cytometry contour plots of IFN-γ + CD8 + T cells among all cell populations in the spleen. (I) Quantification results and (J) representative images of IFN- γ -secreting immune cells in the spleen of mice analyzed by enzyme-linked immunospot (ELISpot) assay. Data were shown as mean ± SD (n = 3).

Article Snippet: Quantification analysis of (C) CD3 + CD4 + T cells and (D) CD3 + CD8 + T cells in the spleen. (E) Quantification analysis and (F) representative flow cytometry contour plots of OVA-specific CD8 + T cells among all cell populations in the spleen. (G) Quantification analysis and (H) representative flow cytometry contour plots of IFN-γ + CD8 + T cells among all cell populations in the spleen. (I) Quantification results and (J) representative images of IFN- γ -secreting immune cells in the spleen of mice analyzed by enzyme-linked immunospot (ELISpot) assay.

Techniques: In Vivo, Activation Assay, Injection, Flow Cytometry, Enzyme-linked Immunospot

Journal: Nucleic Acids Research

Article Title: Transient SUMOylation inhibition in human pre-adipocytes stably imprints a transcriptional beiging fate

doi: 10.1093/nar/gkag232

Figure Lengend Snippet: Stable mobilization of CEBPs and PPARG upon transient TAK-981 treatment in pre-adipocytes. ( A ) ATAC-seq experimental layout in hTERT A41hWAT-SVF pre-adipocytes. Time series analysis and hierarchical clustering of significant variations in chromatin accessibility at day 0 (D0), 12 h (12h), and day (D7) after TAK-981 treatment. ( C ) GOBP analysis of ATAC-seq clusters 2 and 6 revealed in panel (B). See and for an analysis of all clusters. ( D – F ) Volcano plots displaying the results of ATAC-seq inferred differential TF activity analyses performed at D0, 12h, and D7 after TAK-981 or DMSO treatment. Colored dots indicate significant differentially mobilized TFs (TAK-981 versus DMSO). Differential binding score >0.05 and pAdj <0.001. ( G ) Time series analysis inferred TF activity over time (TAK-981 versus DMSO) at D0, 12h, and D7 after TAK-981 treatment. ( H ) Western blot analysis of CEBPB SUMOylation in DMSO and TAK-981-treated cells. ( I ) Western blot analysis of CEBPB and PPARG in DMSO, rosiglitazone, TAK-981-treated cells and cotreated hTERT A41hWAT-SVF pre-adipocytes. ( J ) Western blot analysis of PPARG in DMSO, rosiglitazone, TAK-981-treated cells and cotreated hASCs.

Article Snippet: Samples for assay for transposase-accessible chromatin sequencing (ATAC-seq) were prepared based on previously published protocols with the Illumina ATAC-seq library preparation kit (20034197) [ – ].

Techniques: Activity Assay, Binding Assay, Western Blot