multiplex immunofluorescence staining kit  (Servicebio Inc)

Journal: NPJ Parkinson's Disease

Article Title: LRRK2 R1627P mutation amplifies environmental risk factors induced chronic inflammation and α-synuclein aggregation in the gut of rats

doi: 10.1038/s41531-026-01281-3

Figure Lengend Snippet: a Volcano plot comparing DEGs for RNA-seq in LRRK2 R1627P vs WT, LRRK2 −/− vs WT ( p value < 0.05, fold change >1.2), n = 3. b BP categories in GO enrichment analysis and KEGG pathway of DEGs in LRRK2 R1627P vs WT, n = 3. c BP categories in GO enrichment analysis and KEGG pathway of DEGs in LRRK2 −/− vs WT, n = 3. d Western blot analysis for TLR4, MyD88 and NF-κB protein levels in the small intestine of rats, n = 4. One-way ANOVA was used. e TEM images of rats small intestinal LP. Ec enterocytes, LP lamina propria. Scale bars 5 μm, n = 3. One-way ANOVA was used. f Western blot analysis for CD68 and CD163 protein levels in the small intestine of rats, n = 4. One-way ANOVA was used. g Immunofluorescence staining images of CD68 (green) and CD163 (red) (upper panel), IL-6 (green) and iNOS (red) (down panel) positive cells in the small intestinal LP of rats. Nuclei were stained with DAPI (blue). Scale bar 100 μm, n = 4. One-way ANOVA was used. h Flow cytometry scatter plot of small intestianl LP isolated from rats, n = 3. CD11b + CD68 + CD163 - cells represent M1 population and CD11b + CD68 - CD163 + cells represent M2 population. i Statistical analysis of total macrophages, M1 macrophages and M2 macrophages in the small intestianl LP using flow cytometry. One-way ANOVA was used. j Immunofluorescence staining images of CD68 (green) and p S129 -α-Syn (red) positive cells in the small intestinal LP of rats. Nuclei were stained with DAPI (blue). Scale bar 100 μm, n = 4. k Statistical analysis of immunofluorescence staining. One-way ANOVA was used. Data are presented as mean ± SEM, * p < 0.05, ** p < 0.01.

Article Snippet: Multiplex immunofluorescence staining kit (G1236) was purchased from Servicebio (Wuhan, China).

Techniques: RNA Sequencing, Western Blot, Immunofluorescence, Staining, Flow Cytometry, Isolation

Journal: NPJ Parkinson's Disease

Article Title: LRRK2 R1627P mutation amplifies environmental risk factors induced chronic inflammation and α-synuclein aggregation in the gut of rats

doi: 10.1038/s41531-026-01281-3

Figure Lengend Snippet: a HE staining images of the small intestine of LPS-treated rats. The green circles in the lower panels highlight the red blood cells in the LP. Scale bar 100 μm, n = 4. b Images of goblet cells stained with AB-PAS in the small intestine of LPS-treated rats. Scale bar 100 μm, n = 4. c Immunohistochemical staining images of Cleaved caspase 3 in the small intestine of LPS-treated rats. Arrows show apoptotic IECs. Scale bar 100 μm, n = 4. d Western blot analysis for ZO-1, E-cadherin, TLR4, MyD88, NF-κB, CD68 and CD163 protein levels in the small intestine of LPS-treated rats, n = 4. e Immunofluorescence staining images and quantification of CD68/IL-6 (green) and CD163/iNOS/p S129 -α-Syn (red) positive cells in the small intestinal LP of LPS-treated rats. Nuclei were stained with DAPI (blue). Scale bar 100 μm, n = 4. One-way ANOVA was used. Data are presented as mean ± SEM, * p < 0.05, ** p < 0.01.

Article Snippet: Multiplex immunofluorescence staining kit (G1236) was purchased from Servicebio (Wuhan, China).

Techniques: Staining, Immunohistochemical staining, Western Blot, Immunofluorescence

Journal: NPJ Parkinson's Disease

Article Title: LRRK2 R1627P mutation amplifies environmental risk factors induced chronic inflammation and α-synuclein aggregation in the gut of rats

doi: 10.1038/s41531-026-01281-3

Figure Lengend Snippet: a , b Western blot analysis for TLR4, MyD88, NF-κB, CD68 and CD163 protein levels in the small intestine of WT, LRRK2 R1627P and LRRK2 −/− rats treated with saline (control) or TLR4 inhibitor (TAK-242), n = 4. One-way ANOVA was used. c Immunofluorescence staining images and quantification of CD68/IL-6 (green) and CD163/iNOS/p S129 -α-Syn (red) positive cells in the small intestinal LP of rats treated with saline (control) or TLR4 inhibitor (TAK-242). Nuclei were stained with DAPI (blue). Scale bar 100 μm, n = 4. Two-way ANOVA was used; for CD68/IL-6/iNOS analysis interaction p < 0.01; for CD163 analysis interaction p > 0.05; for p S129 -α-Syn analysis interaction p < 0.001. d GO enrichment analysis for DEGs in ConRP vs ConWT, TAKRP vs ConRP, TAKRP vs ConWT, n = 3. e Heatmap of DEGs from ConWT, ConRP and TAKRP group, n = 3. Data are presented as mean ± SEM, * p < 0.05, ** p < 0.01.

Article Snippet: Multiplex immunofluorescence staining kit (G1236) was purchased from Servicebio (Wuhan, China).

Techniques: Western Blot, Saline, Control, Immunofluorescence, Staining

Journal: eBioMedicine

Article Title: Clonal GZMK + CD8 + T cells are identified as a hallmark of the pathogenesis of cGVHD-induced bronchiolitis obliterans syndrome after allogeneic hematopoietic stem cell transplantation

doi: 10.1016/j.ebiom.2024.105535

Figure Lengend Snippet: Bosutinib inhibits GZMK + CD8 + T cells in vitro and alleviates lung fibrosis in murine cGVHD-BOS mode l . (A–B) Human T cells were incubated with 1ug/ml lipopolysaccharide in the presence of 10 μM Bosutinib or DMSO control for 48 h. The frequencies of GZMK + cells in CD8 + T cells were detected by flow cytometry (n = 4) (A). Western-Blot detected the protein expression levels of phosphorylated-Src, Src and GZMK (n = 5). GAPDH was used as a loading control. Numbers indicate the relative optical density of target protein levels to GAPDH. The relative p-Src/Src and GZMK values were calculated and compared between two groups using a paired T-test (B). (C) Pulmonary function analysis of resistance and compliance of mice in control, cGVHD-BOS group and Bosutinib group on day 56 after transplantation. (D) Body weight, GVHD severity and overall survival rate of mice in without cGVHD group (abbreviated as control here), cGVHD-BOS group and cGVHD-BOS with Bosutinib treatment group (abbreviated as Bosutinib here). The cGVHD-BOS mice were randomly assigned to the treatment group with intraperitoneal injection of Bosutinib as 100 mg/kg every 3 days during days 28–56 post-transplantation. (E) HE (upper) and Masson staining (lower) of lung tissues of mice from control, cGVHD-BOS group and Bosutinib group on day 56 after transplantation. (F) Culture supernatant concentration levels of Granzym K secreted by CD8 + T cells sorted from the lung tissues of mice from control, cGVHD-BOS group and Bosutinib group on day 56 after transplantation. CD8 + T cells were stimulated by PMA and ionomycin for 24 h in vitro . (G) Multiple Immunofluorescence staining of the lung tissues of mice from control, cGVHD-BOS group and Bosutinib group on day 56 after transplantation. (DAPI-blue, CD8-red, GranzymeK-green, CollegenI-pink). Data in C, D, F represent 3 independent experiments (control, n = 8; cGVHD-BOS group, n = 8; Bosutinib group, n = 8). Comparisons of body weight and GVHD severity between two groups were performed using ANOVA test. Difference of overall survival rate between two groups was performed using log-rank test. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

Article Snippet: Multiplexed immunofluorescence staining of the lung tissue was performed using multiplexed immunofluorescence staining kit (GDP1014, Servicebio) according to manufacturer's instruction.

Techniques: In Vitro, Incubation, Control, Flow Cytometry, Western Blot, Expressing, Transplantation Assay, Injection, Staining, Concentration Assay, Immunofluorescence