multiplex immunofluorescence staining  (Akoya Biosciences)

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: Frontiers in Immunology

Article Title: Selective inhibition of canonical STAT3 signaling suppresses K-ras mutant lung tumorigenesis and reinvigorates anti-tumor immunity

doi: 10.3389/fimmu.2025.1575181

Figure Lengend Snippet: STAT3 inhibition increases DC and Th1 proportion within the TIME. (A) Representative dot plots of myeloid cell flow cytometry analysis pre-gated on CD11c + cells; expression of Ly6C and CD11b is used to delineate classical type 1 DCs (cDC1s)/alveolar macrophages (AMs), cDC2s, and monocytic DCs (Mo-DCs), and is quantified in (B) as a percentage of CD45 + cells (N = 3-5). (C) Representative dot plots of lymphoid cell flow cytometry analysis with ex vivo restimulation by PMA/Ionomycin, pre-gated on CD3 + cells; expression of CD4 and IFNγ is used to mark T helper 1 (Th1) cells and is quantified in (D) as a percentage of CD4 + cells (N = 3-5). (E) Representative multiplex immunofluorescence (COMET) images for B cells (B220 + ); original magnification, 20X; scale bar is 100 µm. Data represent mean ± SEM. **P < 0.01, *P < 0.5 by unpaired t test.

Article Snippet: Multiplex immunofluorescence staining (Lunaphore COMET) was performed on FFPE-derived sections as described in Wang et al. ( ).

Techniques: Inhibition, Flow Cytometry, Expressing, Ex Vivo, Multiplex Assay, Immunofluorescence

Journal: iScience

Article Title: Osteoclast-derived arachidonic acid triggers dormant lung adenocarcinoma cell activation

doi: 10.1016/j.isci.2025.112167

Figure Lengend Snippet: CD36-dependent lipid uptake drives activation of dormant A549 cells (A) IF comparison of intracellular lipid content in dormant A549 cells after treatment with OC-CM and OC-CM combined with SSO (scale bars: 25 μm). (B) Quantification of lipid content in (A). (C) IF comparison of Ki-67 expression in dormant A549 cells after treatment with OC-CM and OC-CM combined with SSO (scale bars: 25 μm). (D) Quantification of Ki-67 expression in (C). (E) WB comparison of the expression of ANGPTL4 and dormancy markers in A549 cells after co-treatment with the CD36 inhibitor SSO and OC-CM. (F) Quantification of ANGPTL4 and dormancy marker expression in (E). NC: normal control; 3D: Matrigel culture-induced dormancy; DMSO: DMSO treatment for 24 h; OC: osteoclast CM treatment for 24 h; SSO: 50 nM SSO treatment for 24 h (the statistical data are presented as mean ± SD. (F) was analyzed using two-way ANOVA, (B) and (D) using one-way ANOVA, and (L) with a two-tailed t test, with multiple comparisons adjusted by Tukey multiple comparisons test. ns: not significant, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Also see Figure S3 .

Article Snippet: CD36 for Tissue Multiplex Immunofluorescence Staining , WANLEIBIO , Cat# WL02390; RRID: AB_3668969.

Techniques: Activation Assay, Comparison, Expressing, Marker, Control, Two Tailed Test

Journal: iScience

Article Title: Osteoclast-derived arachidonic acid triggers dormant lung adenocarcinoma cell activation

doi: 10.1016/j.isci.2025.112167

Figure Lengend Snippet: AA from osteoclasts triggers dormant A549 cell activation in a dose-dependent manner (A) WB analysis of the activation effect of different concentrations of AA (0.625–5 μM) on dormant A549 cells. (B) Quantification of CD36, ANGPTL4, and dormancy marker expression in (B). (C) ELISA detection of AA content in the CM of RAW264.7 cells and mature osteoclasts. (D) WB analysis of the activation effect of OC-CM on dormant A549 cells after treatment with BW 755C. (E) Quantification of ANGPTL4 and dormancy marker expression in (D). 3D: Matrigel culture-induced dormancy; DMSO: DMSO treatment for 24 h; OC: Osteoclast CM treatment for 24h; AA: 0.625-5 Μm AA treatment for 24 h (The statistical data are presented as mean ± SD. (B) and (E) were analyzed using two-way ANOVA, and (C) with a two-tailed t test, with multiple comparisons adjusted by Tukey multiple comparisons test. ns: not significant, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Also see Figure S4 .

Article Snippet: CD36 for Tissue Multiplex Immunofluorescence Staining , WANLEIBIO , Cat# WL02390; RRID: AB_3668969.

Techniques: Activation Assay, Marker, Expressing, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: iScience

Article Title: Osteoclast-derived arachidonic acid triggers dormant lung adenocarcinoma cell activation

doi: 10.1016/j.isci.2025.112167

Figure Lengend Snippet: Upregulation of CD36 and ANGPTL4 is associated with osteoclast-mediated activation of dormant LUAD cells in vivo (A) Schematic diagram of the bone metastasis mouse model. (B) H&E staining and multiplex immunofluorescence detection of Ki-67, TRAP, CK7, ANGPTL4, and CD36 expression in the tibia and femur of mice without metastasis and with bone metastasis (arrows indicate osteoclasts or Ki-67+ cells). a. Dormant lesions (low Ki-67); b. Activated lesions (high Ki-67) (scale bar: 100 μm). (C) Proportion of Ki-67 positive cells in dormant and activated metastatic lesions. (D) Average fluorescence intensity of TRAP in dormant and activated metastatic lesions. (E) Average fluorescence intensity of ANGPTL4 in dormant and activated metastatic lesions. (F) Average fluorescence intensity of CD36 in dormant and activated metastatic lesions (The statistical data are presented as mean ± SD. (C)–(F) were analyzed using two-tailed t test. ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001.

Article Snippet: CD36 for Tissue Multiplex Immunofluorescence Staining , WANLEIBIO , Cat# WL02390; RRID: AB_3668969.

Techniques: Activation Assay, In Vivo, Staining, Multiplex Assay, Immunofluorescence, Expressing, Fluorescence, Two Tailed Test

Journal: iScience

Article Title: Osteoclast-derived arachidonic acid triggers dormant lung adenocarcinoma cell activation

doi: 10.1016/j.isci.2025.112167

Figure Lengend Snippet:

Article Snippet: CD36 for Tissue Multiplex Immunofluorescence Staining , WANLEIBIO , Cat# WL02390; RRID: AB_3668969.

Techniques: Western Blot, Immunofluorescence, Staining, Multiplex Assay, Recombinant, Enzyme-linked Immunosorbent Assay, shRNA, Sequencing, Software, Gene Expression