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: Journal of Advanced Research

Article Title: CCR1 hi /CCL5 hi macrophage-mediated CCL5 hi T cell chemotaxis in salivary gland aggravates Sjögren’s syndrome

doi: 10.1016/j.jare.2025.06.076

Figure Lengend Snippet: CCR1-overexpressing macrophages induce T and B cell migration. (A) Immunofluorescence (IF) detection of CD68 expression in THP-1 cells induced by PMA (100 nM, 24 h). (B) Quantitative analysis of CD68 expression based on IF staining fluorescence intensity. (C) Schematic diagram of the detection of macrophage-induced T and B cell migration. (D, E) Western blot analysis and quantitative evaluation of CCR1 and CCL5 expression in macrophages after transfection with the CCR1 overexpression plasmid. (F) ELISA measurement of CCL5 concentration in supernatants from CCR1-overexpressing macrophages. (G, H) Transwell assay evaluating macrophage-mediated migration of T and B cells, with quantitative analysis of migrated cell numbers. Statistical significance was determined using one-way ANOVA.

Article Snippet: The release level of CCL5 in cell culture supernatants was assessed using a commercial ELISA kit (EK1129, Lianke Biotechnology Co., Ltd., Hangzhou, China) following the manufacturer’s instructions.

Techniques: Migration, Immunofluorescence, Expressing, Staining, Fluorescence, Western Blot, Transfection, Over Expression, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Concentration Assay, Transwell Assay

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: 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.

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