Journal: bioRxiv

Article Title: Corilagin controls post-parasiticide schistosome egg-induced liver fibrosis by inhibiting Stat6 signalling pathway

doi: 10.1101/340299

Figure Lengend Snippet: Effect of Corilagin on the expression of ARG1 according to ELISA. Data shown are the mean ± SD from 3 independent experiments. # P< 0.05 vs . normal group; *P< 0.05 VS model group; as determined by Student’s t-test; ## P< 0.01 determined by One-way ANOVA.

Article Snippet: The Mouse ARG1, Fizz1, Ym1, TGFβ and PDGF ELISA kits were purchased from Elabscience Biotechnology Co., Ltd (Wuhan, China).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Corilagin controls post-parasiticide schistosome egg-induced liver fibrosis by inhibiting Stat6 signalling pathway

doi: 10.1101/340299

Figure Lengend Snippet: Effect of Corilagin on expression of PDGF, Fizz1, Ym1, and TGFβ according to ELISA. Data shown are the mean ± SD from 6 experiment mice. # P< 0.05, ## P< 0.01 vs . normal group; *P< 0.05, ** P< 0.01 VS model group; As determined by Student’s t-test; ### P< 0.01 determined by One-way ANOVA.

Article Snippet: The Mouse ARG1, Fizz1, Ym1, TGFβ and PDGF ELISA kits were purchased from Elabscience Biotechnology Co., Ltd (Wuhan, China).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Materials Today Bio

Article Title: 3D-printed PRP-infused double-network hydrogels orchestrate inflammation resolution and vascular regeneration in infected wounds

doi: 10.1016/j.mtbio.2026.102841

Figure Lengend Snippet: In vitro Anti-inflammatory ability of hydrogels by driving macrophage polarization. (A) Immunofluorescence images of the M1 and M2-type macrophages in different groups. (B, C) Quantitative analysis of positive cells (CD86 and CD206) fluorescence intensity values. (D) Western blot analysis for the expression of inflammation proteins (Arg-1 and iNOS) in RAW264.7 with hydrogels treatments, and (E, F) Semiquantitative analysis of corresponding protein expression levels. (G–I) ELISA analysis of specific markers of M1 macrophage and M2 macrophage with various treatments. (mean ± SD, n = 3, ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001).

Article Snippet: The amount of growth factors released from the hydrogel was quantified using the QuantiCyto® Rat EGF ELISA Kit (Neobioscience Technology Co, Ltd.), Rat PDGF-A ELISA kit, and Rat VEGF ELISA Kit (CUSABIO, https://www.cusabio.com/ ).

Techniques: In Vitro, Immunofluorescence, Fluorescence, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Cancers

Article Title: Cancer-Associated Fibroblasts Exposed to High-Dose Ionizing Radiation Promote M2 Polarization of Macrophages, Which Induce Radiosensitivity in Cervical Cancer

doi: 10.3390/cancers15051620

Figure Lengend Snippet: CCL2 is critical for high-dose irradiated CAFs to promote M2 polarization. ( A ) mCAFs were exposed to 8 Gy radiation or not (control), and the supernatant of mCAFs was subjected to multiplex cytokine array analysis 24 h later. Left: a representative blot. Right: quantification of dot intensity of significantly changed cytokines. ( B ) Cervical cancer cell lines and CAFs were irradiated with the indicated doses. After 24 h, CCL2 expression was analyzed by Western blot analysis. ( C ) mCAFs and hCAFs were exposed to the indicated doses of radiation. After 24 h, the supernatant was subjected to ELISA. Data are shown as mean ± SD from three independent experiments. The data were analyzed using one-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001. ( D ) BMDMs were treated with 20 ng/mL CCL2 for 24 h and then subjected to flow cytometry analysis. Data are shown as mean ± SD from three independent experiments. Differences between groups were analyzed using unpaired Student’s t -test. * p < 0.05. ( E ) M0 BMDMs treated with 10 nM CCR2 antagonist (INCB3344) were co-cultured with irradiated or non-irradiated mCAFs for 3 days. Then, the cells were subjected to flow cytometry analysis to evaluate CD206 expression. Data are shown as mean ± SD from three independent experiments. * p < 0.05. ( F ) Anti-CCL2 neutralizing antibody (20 μg/mL) was added to a co-culture system consisting of 8 Gy irradiated mCAFs and M0 BMDMs. The BMDMs were subjected to flow cytometry analysis after 3 days of co-cultivation. Data are shown as mean ± SD from three independent experiments. The data were analyzed using one-way ANOVA. * p < 0.05. ( G ) M0 BMDMs were co-cultured with mCAFs exposed to the indicated doses of radiation for 3 days and 10 nM CCR2 antagonist (INCB3344) was added into the 8 Gy irradiated mCAF co-culture system. Immunofluorescence analysis of the expression of Ym-1 (red) in BMDMs. The whole western blots are shown in File S1.

Article Snippet: The supernatant of CAFs and cervical cancer cells was assayed for cytokines and chemokines using the Mouse XL Cytokine Array (Ary028; R&D Systems) and CCL2 ELISA kits (mouse, EK0568; human, EK0441; BOSTER) according to the manufacturers’ instructions.

Techniques: Irradiation, Control, Multiplex Assay, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Cell Culture, Co-Culture Assay, Immunofluorescence

Journal: Cancers

Article Title: Cancer-Associated Fibroblasts Exposed to High-Dose Ionizing Radiation Promote M2 Polarization of Macrophages, Which Induce Radiosensitivity in Cervical Cancer

doi: 10.3390/cancers15051620

Figure Lengend Snippet: Model summarizing the proposed signaling events among CAFs, TAMs, and cervical cancer cells under high-dose radiation. CAFs secrete an elevated level of CCL2 upon treatment with high-dose radiation. CCL2 promotes pro-tumor transition in macrophages. M2 macrophages induce radioresistance in cervical cancer cells.

Article Snippet: The supernatant of CAFs and cervical cancer cells was assayed for cytokines and chemokines using the Mouse XL Cytokine Array (Ary028; R&D Systems) and CCL2 ELISA kits (mouse, EK0568; human, EK0441; BOSTER) according to the manufacturers’ instructions.

Techniques:

Journal: Scientific Reports

Article Title: Single-nucleus transcriptomic profiling of the diaphragm during mechanical ventilation

doi: 10.1038/s41598-024-82530-4

Figure Lengend Snippet: Differentially expressed genes and Kyoto Encyclopedia of Genes and Genomes analysis. Volcano plots showing the top twenty up- or downregulated genes for each cell type. The red dots represent upregulated genes, and the blue dots represent downregulated genes. ( P value < 0.05, and |log2foldchange| > 0.58). Bubble plot indicating the top enriched pathways for each cell type based on KEGGpathway enrichment analysis of differentially expressed genes( www.kegg.jp/kegg/kegg1.html ). The sizes of the dots represent the number of genes included in each pathway. The colour gradient of dots represents the adjusted P values of each enriched pathway. The genes Pfkfb3 , Tbc1d1 and the insulin signaling pathway; Pdgfd and the PI3K-Akt signaling pathway; Cxcr2 and PLD, Rap1 signaling pathways; Ccl21 and chemical carcinogenesis-reactive oxygen species signaling pathway; Mef2c and the calcium signaling pathway; Negr1 and leukocyte transendothelial migration signaling pathway are labelled by the black and red bars in Panels a , b , c , d , e , and f .

Article Snippet: After the semidry blotting procedure (50 min, 90 V), the membrane was incubated for 1 h at room temperature (RT) in 5% BSA blocking solution, followed by overnight incubation on a shaker at 4 °C with primary antibodies against PFKFB3 (bs-3528R, Boster Biological Technology Co., Ltd., Wuhan, China), PDGFD (bs-24572R, Boster), CXCR2 (bs-1629R, Boster), NEGR1 (bs-11095R, Boster), SEMA3A (bs-10468R, Boster), MEF2C (bs-4130R, Boster) and β-actin (66009-1-Ig, Proteintech, Wuhan, China) as a loading control.

Techniques: Protein-Protein interactions, Migration

Journal: Scientific Reports

Article Title: Single-nucleus transcriptomic profiling of the diaphragm during mechanical ventilation

doi: 10.1038/s41598-024-82530-4

Figure Lengend Snippet: High-dimensional weighted gene coexpression network analysis (hdWGCNA). a The top left panel depicts the soft power threshold for choosing a scale-free topology model. The average connectivity of the topological network was most stable at the lowest soft threshold of 6. b Dynamic Tree Cut algorithm for gene clustering. Each leaf on the tree represents a gene, and the colour at the bottom indicates the assignment to a specific coexpression module. c Bubble plots displaying the scores obtained for eight modules in eight cell subtypes. d snRNA-seq UMAP coloured by module eigengene (ME) for eight coexpression modules. e The eight modules’ values of -log10 (Adj P value) on the Y-axis versus average log2 (fold change) on the X-axis are shown for each cell type, including endothelial cells, fibroblast 1, fibroblast 2, myocytes, neurocytes, pericytes, satellite cells and smooth muscle cells. The larger the value is, the greater the fit between the coloured module and the specific cell type. The plot shows that fibroblast 1 features a brown module, fibroblast 2 features a green module, endothelial cells feature a yellow module, and myocyte cells feature a turquoise module. f UMAP plot of the gene coexpression network with eight coloured modules and the top two hub genes labelled in each module. Nodes are coloured according to coexpression module assignment. g Hub genes in each module were identified by eigengene-based connectivity (KME). The following five hub genes associated with fibrosis were identified: Pfkfb3 and Tbc1d1 in the brown module, Pdgfd in the green module, Sema3a in the yellow module and Ryr3 in the turquoise module.

Article Snippet: After the semidry blotting procedure (50 min, 90 V), the membrane was incubated for 1 h at room temperature (RT) in 5% BSA blocking solution, followed by overnight incubation on a shaker at 4 °C with primary antibodies against PFKFB3 (bs-3528R, Boster Biological Technology Co., Ltd., Wuhan, China), PDGFD (bs-24572R, Boster), CXCR2 (bs-1629R, Boster), NEGR1 (bs-11095R, Boster), SEMA3A (bs-10468R, Boster), MEF2C (bs-4130R, Boster) and β-actin (66009-1-Ig, Proteintech, Wuhan, China) as a loading control.

Techniques:

Journal: Scientific Reports

Article Title: Single-nucleus transcriptomic profiling of the diaphragm during mechanical ventilation

doi: 10.1038/s41598-024-82530-4

Figure Lengend Snippet: Protein-protein interaction network (PPI network).We constructed nine PPI networks displaying protein-protein interactions among the related genes. The nodes represent proteins, and the edges represent the interaction strength between two proteins. The proteins PFKFB3 ( a ), PDGFD ( b ), CXCR2 ( c ), CCL21 ( d ), SEMA3A ( e ), RYR3 ( f ), MEF2C ( g ), NEGR1( h ) and TBC1D1 ( i ) which are located at the hub of the interaction network, are responsible for diaphragm fibrosis and atrophy.

Article Snippet: After the semidry blotting procedure (50 min, 90 V), the membrane was incubated for 1 h at room temperature (RT) in 5% BSA blocking solution, followed by overnight incubation on a shaker at 4 °C with primary antibodies against PFKFB3 (bs-3528R, Boster Biological Technology Co., Ltd., Wuhan, China), PDGFD (bs-24572R, Boster), CXCR2 (bs-1629R, Boster), NEGR1 (bs-11095R, Boster), SEMA3A (bs-10468R, Boster), MEF2C (bs-4130R, Boster) and β-actin (66009-1-Ig, Proteintech, Wuhan, China) as a loading control.

Techniques: Construct, Protein-Protein interactions

Journal: Scientific Reports

Article Title: Single-nucleus transcriptomic profiling of the diaphragm during mechanical ventilation

doi: 10.1038/s41598-024-82530-4

Figure Lengend Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. qRT-PCR and Western blotting showing the expression of selected genes and corresponding proteins in the mechanically ventilated diaphragm and control ones. The bar graphs show the quantification of the mRNA expression of Pfkfb3 ( a ), Pdgfd ( b ), Cxcr2 ( c ), Negr1 ( d ), Sema3a ( e ), and Mef2c ( f ) normalized to that of GAPDH. Asterisks indicate significant differences ( n = 3 in each group) (* P < 0.05). Western blotting analysis of the protein expression of PFKFB3 ( g ), PDGFD ( h ), CXCR2 ( i ), NEGR1 ( j ), SEMA3A ( k ), and MEF2C ( l ) normalized to that of β-actin as a loading control.

Article Snippet: After the semidry blotting procedure (50 min, 90 V), the membrane was incubated for 1 h at room temperature (RT) in 5% BSA blocking solution, followed by overnight incubation on a shaker at 4 °C with primary antibodies against PFKFB3 (bs-3528R, Boster Biological Technology Co., Ltd., Wuhan, China), PDGFD (bs-24572R, Boster), CXCR2 (bs-1629R, Boster), NEGR1 (bs-11095R, Boster), SEMA3A (bs-10468R, Boster), MEF2C (bs-4130R, Boster) and β-actin (66009-1-Ig, Proteintech, Wuhan, China) as a loading control.

Techniques: Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Expressing, Control