Journal: PLoS Biology

Article Title: Improved genetically encoded near-infrared fluorescent calcium ion indicators for in vivo imaging

doi: 10.1371/journal.pbio.3000965

Figure Lengend Snippet: ( a ) Mutations of NIR-GECO2 and NIR-GECO2G relative to NIRGECO1. The different mutations between NIR-GECO2 and NIR-GECO2G are highlighted in green. ( b ) Relative fluorescence intensity (mean ± SEM) of NIR-GECO1, NIR-GECO2, NIR-GECO2G, and miRFP720 in neurons ( n = 160, 120, 219, and 84 neurons, respectively, from 2 cultures). Fluorescence was normalized by co-expression of GFP via self-cleavable 2A peptide. ( c – e ) Comparison of NIR-GECO variants, as a function of stimulus strength (the same color code is used in panels c – e ). ( c ) ΔF/F 0 ; ( d ) rise time; ( e ) half decay time. Values are shown as mean ± SD ( n = 10 wells from 3 cultures). The underlying data for ( b ) to ( e ) can be found in . GFP, green fluorescent protein; NIR, near-infrared; SD, standard deviation; SEM, standard error of the mean.

Article Snippet: Plasmids pAAV-CAG-NIR-GECO2 (plasmid no. 159603), pAAV-NIR-GECO2G (plasmid no. 159605), and pSF11-wNIR-GECO2-T2A-HO1(plasmid no.159606) are available via Addgene according to the terms of the Uniform Biological Material Transfer Agreement.

Techniques: Fluorescence, Expressing, Comparison, Standard Deviation

Journal: PLoS Biology

Article Title: Improved genetically encoded near-infrared fluorescent calcium ion indicators for in vivo imaging

doi: 10.1371/journal.pbio.3000965

Figure Lengend Snippet: ( a – c ) Fluorescence traces of NIR-GECO2G, NIR-GECO2, and NIR-GECO1 in response to 100 ms ( a ), 500 ms ( b ), and 1 s ( c ) blue light activation (470 nm at a power of 1.9 mW/mm 2 ) in HeLa cells with co-expression of Opto-CRAC. Opto-CRAC is composed of the STIM1-CT and LOV2 domain. The fusion of STIM1-CT to the LOV2 domain allows photo-controllable exposure of the active site of SRIMI-CT, which is able to interact with ORAI1 and trigger Ca 2+ entries across the plasma membrane . Black, green, and dark blue lines represent averaged data for NIR-GECO2G ( n = 32 cells), NIR-GECO2 ( n = 25 cells), and NIR-GECO1 ( n = 23 cells), respectively. The same color code is used in panels a–c . Shaded areas represent the SD. ( d ) Quantitative -ΔF/F 0 (mean ± SD) for NIR-GECO2G, NIR-GECO2, and NIR-GECO1 in a–c . The underlying data for a – d can be found in . NIR, near-infrared; SD, standard deviation.

Article Snippet: Plasmids pAAV-CAG-NIR-GECO2 (plasmid no. 159603), pAAV-NIR-GECO2G (plasmid no. 159605), and pSF11-wNIR-GECO2-T2A-HO1(plasmid no.159606) are available via Addgene according to the terms of the Uniform Biological Material Transfer Agreement.

Techniques: Fluorescence, Activation Assay, Expressing, Clinical Proteomics, Membrane, Standard Deviation

Journal: PLoS Biology

Article Title: Improved genetically encoded near-infrared fluorescent calcium ion indicators for in vivo imaging

doi: 10.1371/journal.pbio.3000965

Figure Lengend Snippet: ( a ) Left, fluorescent image of neurons expressing NLS-jGCaMP7s (λ ex = 488-nm laser light, λ em = 527/50 nm). Right, fluorescent image of neurons expressing NIR-GECO2-T2A-HO1 (λ ex = 640-nm laser light, λ em = 685/40 nm). Representative of more than 3 worms, both under tag-168 promoter. Scale bar, 50 μm. ( b ) Fluorescence traces of NLS-jGCaMP7s (top) and NLS-NIR-GECO2 (bottom) in response to the stimulation of microfluidic containing 200 mM NaCl. Solid lines represent averaged data from 3 neurons. Shaded areas are shown as SD. Triangles on the top of the traces indicate the time points of stimulation (20 seconds for each stimulation). ( c ) Quantitative fluorescence changes of NLS-jGCaMP7s and NLS-NIR-GECO2 in b ( n = 36 spikes from 3 neurons). ( d ) Fluorescence image of the 4 C . elegans expressing NIR-GECO2-T2A-HO1 in AVA neurons (under flp-18 promoter) and CoChR-GFP in ASH neurons (under sra-6 promoter). The merged image is shown. Imaging conditions: NIR-GECO2, λ ex = 640-nm laser light, λ em = 685/40; GFP, λ ex = 488-nm laser light, λ ex = 527/50 nm. ( e ) Individual traces of NIR-GECO2 fluorescence in an AVA neuron under blue light illumination (20 mW/mm 2 , λ ex = 488-nm laser light, 100 ms; blue bars). The underlying data for b , c , and e can be found in . NIR, near-infrared; SD, standard deviation.

Article Snippet: Plasmids pAAV-CAG-NIR-GECO2 (plasmid no. 159603), pAAV-NIR-GECO2G (plasmid no. 159605), and pSF11-wNIR-GECO2-T2A-HO1(plasmid no.159606) are available via Addgene according to the terms of the Uniform Biological Material Transfer Agreement.

Techniques: Expressing, Fluorescence, Imaging, Standard Deviation

Journal: PLoS Biology

Article Title: Improved genetically encoded near-infrared fluorescent calcium ion indicators for in vivo imaging

doi: 10.1371/journal.pbio.3000965

Figure Lengend Snippet: ( a ) Fluorescence intensity of NLS-jGCaMP7s and NIR-GECO2 in C . elegans neurons at resting state. Fluorescence was normalized to the same excitation intensity ( n = 132 ROIs from 5 worms; data are shown as mean ± SD) ( b ) SBR of NLS-jGCaMP7s and NIR-GECO2 in neurons of C . elegans at resting state ( n = 132 ROIs from 5 worms; data are shown as mean ± SD). SBR was obtained via dividing the fluorescence intensity from neurons by the averaged autofluorescence from the intestine area. ( c ) SNR of NLS-jGCaMP7s and NLS-NIR-GECO2 quantified from spontaneously spiking neurons (n = 78 ROIs from 4 worms; data are shown as mean ± SD). SNR was calculated by dividing the fluorescence change associated with a spike by the SD of the baseline fluorescence over the 2-second period immediately before the spike. ( d ) The ratio of SBR NIR-GECO2 to SBR NLS-jGCaMP7s at different imaging depths ( n = 5 worms; data are shown as mean ± SD). ( e ) The ratio of SNR NLS-NIR-GECO2 to SNR NLS-jGCaMP7s at different imaging depths ( n = 4 worms; data are shown as mean ± SD). NIR-GECO2 (without NLS) and NLS-jGCaMP7s were used for the experiments in a , b , and d ; NLS-NIR-GECO2 and NLS-jGCaMP7s were used for the experiments in c and e . The underlying data for a – e can be found in . ROI, region of interest; SBR, signal-to-background ratio; SD, standard deviation; SNR, signal-to-noise ratio.

Article Snippet: Plasmids pAAV-CAG-NIR-GECO2 (plasmid no. 159603), pAAV-NIR-GECO2G (plasmid no. 159605), and pSF11-wNIR-GECO2-T2A-HO1(plasmid no.159606) are available via Addgene according to the terms of the Uniform Biological Material Transfer Agreement.

Techniques: Fluorescence, Imaging, Standard Deviation

Journal: Frontiers in Microbiology

Article Title: Akkermansia muciniphila alleviates antibiotic- and LPS-induced oxidative stress via the p38α MAPK–Nrf2 signaling axis

doi: 10.3389/fmicb.2026.1753421

Figure Lengend Snippet: AKK alleviates ABX/LPS-induced pathological remodeling and inflammation in murine intestine via inhibition of the p38α MAPK pathway. (A) Schematic of the experimental design, illustrating the timeline of ABX/LPS challenge and the interventions with AKK or the p38α MAPK inhibitor (SB203580). (B) Body weight changes of mice in different treatment groups. (C) Disease activity index. The data points represent the daily measurements. To make the graph clear, the data at key time points are shown in the figure. (D) Representative images showing: (i) The gross appearance of the colon and cecum; (ii) H&E-stained sections of the ileum; and (iii) H&E-stained sections of the colon. (E) Quantitative analysis of colon length. (F) Histopathological colitis scores. (G–I) Morphometric analysis of the ileum, including villus height, crypt depth, and the villus height-to-crypt depth ratio (VCR). n = 6 independent biological replicates. All data are presented as mean ± SEM. # p < 0.05, ### p < 0.001, #### p < 0.0001 versus NC group; * p < 0.05, ** p > p < 0.01, *** p < 0.001, **** p < 0.0001 versus NS + ABX/LPS group, + p < 0.05, ++ p < 0.01, +++ p < 0.01, +++ p < 0.001, +++p < 0.0001 versus the AKK-treated group.

Article Snippet: Antibodies against p-p38α MAPK (#4511), HO-1(#43966), NQO-1(#3187) were purchased from Cell Signaling Technology (Danvers, MA, United States).

Techniques: Inhibition, Activity Assay, Staining

Journal: Frontiers in Microbiology

Article Title: Akkermansia muciniphila alleviates antibiotic- and LPS-induced oxidative stress via the p38α MAPK–Nrf2 signaling axis

doi: 10.3389/fmicb.2026.1753421

Figure Lengend Snippet: AKK directly inhibits phosphorylation and activation of p38α MAPK in human colonic epithelial cells. In the LPS-induced injury model of Ncm460 cells: (A) Identification of the optimal concentration and treatment time for AKK via CCK-8 assay. (B) AKK reverses the LPS-induced suppression of cell viability. Data are normalized to the control group (set as 1.0). (C) Representative western blot images of p38α MAPK and phosphorylated p38α MAPK (p-p38α MAPK). Blots for p38α MAPK and p-p38α MAPK were obtained from the same membrane. (D) Quantitative analysis of total p-p38α MAPK protein expression levels. n = 6 independent biological replicates. All data are presented as mean ± SEM. # p < 0.05, ### p < 0.001, #### p < 0.0001 versus p38KD group. * p < 0.05, ** p > p < 0.01, *** p < 0.001, **** p < 0.0001 versus LPS group.

Article Snippet: Antibodies against p-p38α MAPK (#4511), HO-1(#43966), NQO-1(#3187) were purchased from Cell Signaling Technology (Danvers, MA, United States).

Techniques: Phospho-proteomics, Activation Assay, Concentration Assay, CCK-8 Assay, Control, Western Blot, Membrane, Expressing

Journal: Frontiers in Microbiology

Article Title: Akkermansia muciniphila alleviates antibiotic- and LPS-induced oxidative stress via the p38α MAPK–Nrf2 signaling axis

doi: 10.3389/fmicb.2026.1753421

Figure Lengend Snippet: Inhibition of p38α MAPK alleviates oxidative stress in human colonic epithelial cells by activating the Nrf2 pathway. In the LPS-induced injury model of Ncm460 cells: (A,B) Representative flow cytometry scatter plots and quantitative analysis of intracellular ROS levels. (C) Levels of cellular oxidative stress markers (GSH, SOD, MDA). (D) Representative western blot images of Nrf2 and its downstream targets HO-1 and NQO1. The representative western blot images presented here are accompanied by the corresponding internal control detected on the same membrane. (E–G) Quantitative analysis of protein and mRNA expression levels of Nrf2 (E) , HO-1 (F) , and NQO1 (G) . n = 6 independent biological replicates. All data are presented as mean ± SEM. # p < 0.05, ### p < 0.001, #### p < 0.0001 versus p38KD group. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus LPS group.

Article Snippet: Antibodies against p-p38α MAPK (#4511), HO-1(#43966), NQO-1(#3187) were purchased from Cell Signaling Technology (Danvers, MA, United States).

Techniques: Inhibition, Flow Cytometry, Western Blot, Control, Membrane, Expressing

Journal: Frontiers in Microbiology

Article Title: Akkermansia muciniphila alleviates antibiotic- and LPS-induced oxidative stress via the p38α MAPK–Nrf2 signaling axis

doi: 10.3389/fmicb.2026.1753421

Figure Lengend Snippet: AKK enhances migration and tight junction integrity in human colonic epithelial cells in a p38α MAPK-dependent manner. In the LPS-induced injury model of Ncm460 cells: (A) Representative images and quantitative analysis of wound closure rate in the scratch assay. (B) Representative western blot images of tight junction proteins. The representative western blot images presented here are accompanied by the corresponding internal control detected on the same membrane. (C–E) Quantitative analysis of protein and mRNA expression levels of ZO-1 (C) , Occludin (D) , and Claudin-1 (E) . n = 6 independent biological replicates. # p < 0.05, ### p < 0.001, #### p < 0.0001 versus p38KD group. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus LPS group.

Article Snippet: Antibodies against p-p38α MAPK (#4511), HO-1(#43966), NQO-1(#3187) were purchased from Cell Signaling Technology (Danvers, MA, United States).

Techniques: Migration, Wound Healing Assay, Western Blot, Control, Membrane, Expressing

Journal: Frontiers in Microbiology

Article Title: Akkermansia muciniphila alleviates antibiotic- and LPS-induced oxidative stress via the p38α MAPK–Nrf2 signaling axis

doi: 10.3389/fmicb.2026.1753421

Figure Lengend Snippet: Akk restores the intestinal barrier by upregulating MUC2 and tight junction expression in a p38α MAPK-dependent manner. (A) Modulation of the mucus barrier by Akk and the p38α MAPK inhibitor (SB203580): representative IHC images of MUC2 and quantitative analysis. (B) Representative western blot images of tight junction proteins across groups. The representative western blot images presented here are accompanied by the corresponding internal control detected on the same membrane. (C–E) Quantitative analysis of protein and mRNA expression levels of ZO-1 (C) , occludin (D) , and claudin-1 (E) . (F,G) Serum levels of intestinal barrier damage markers, DAO activity, and D-LA concentration. n = 6 independent biological replicates. All data are presented as mean ± SEM. # p < 0.05, ### p < 0.001, #### p < 0.0001 versus NC group; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus NS + ABX/LPS group, + p < 0.05, ++ p < 0.01, +++ p < 0.01, +++ p < 0.001, +++ p < 0.0001 versus the AKK-treated group.

Article Snippet: Antibodies against p-p38α MAPK (#4511), HO-1(#43966), NQO-1(#3187) were purchased from Cell Signaling Technology (Danvers, MA, United States).

Techniques: Expressing, Western Blot, Control, Membrane, Activity Assay, Concentration Assay

Journal: Frontiers in Microbiology

Article Title: Akkermansia muciniphila alleviates antibiotic- and LPS-induced oxidative stress via the p38α MAPK–Nrf2 signaling axis

doi: 10.3389/fmicb.2026.1753421

Figure Lengend Snippet: AKK alleviates intestinal oxidative stress by activating the Nrf2 signaling pathway through inhibition of p38α MAPK. (A) Levels of oxidative stress markers in colonic tissue: GSH, SOD, and MDA (B) Representative western blot images of Nrf2 and its downstream targets, HO-1 and NQO1. The representative western blot images presented here are accompanied by the corresponding internal control detected on the same membrane. n = 6 independent biological replicates. (C–E) Quantitative analysis of protein and mRNA expression levels of Nrf2 (C) , HO-1 (D) , and NQO1 (E) . n = 6 independent biological replicates. All data are presented as mean ± SEM. # p < 0.05, ### p < 0.001, #### p < 0.0001 versus NC group; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus NS + ABX/LPS group, + p < 0.05, ++ p < 0.01, +++ p < 0.01, +++ p < 0.001, +++ p < 0.0001 versus the A-treated group.

Article Snippet: Antibodies against p-p38α MAPK (#4511), HO-1(#43966), NQO-1(#3187) were purchased from Cell Signaling Technology (Danvers, MA, United States).

Techniques: Inhibition, Western Blot, Control, Membrane, Expressing

Journal: Frontiers in Microbiology

Article Title: Akkermansia muciniphila alleviates antibiotic- and LPS-induced oxidative stress via the p38α MAPK–Nrf2 signaling axis

doi: 10.3389/fmicb.2026.1753421

Figure Lengend Snippet: Molecular docking predicts direct binding of AKK to p38α MAPK and validation in a murine model. Exploratory molecular docking results demonstrating strong binding between AKK and p38α MAPK: (A) Detailed view of the binding mode, with a binding free energy of −7.0 kcal·mol −1 ; (B) 3D representation of binding site interactions (yellow dashed lines indicate hydrogen bonds); (C) Schematic 3D structure of the complex; (D) 2D diagram of binding site interactions. (E) Top ten ranked binding conformations from docking screening. (F–I) Experimental validation in an ABX/LPS-induced mouse intestinal injury model: (F) Representative western blot images of p38α MAPK and p-p38α MAPK in colonic tissue; Blots for p38α MAPK and p-p38α MAPK were obtained from the same membrane. (G) p38α MAPK mRNA expression levels in the colon; (H) p-p38α MAPK/p38α MAPK ratio; (I) Total p-p38α MAPK protein levels. n = 6 independent biological replicates. All data are presented as mean ± SEM. # p < 0.05, ### p < 0.001, #### p < 0.0001 versus NC group; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus NS + ABX/LPS group, + p < 0.05, ++ p < 0.01, +++ < 0.01, +++ p < 0.001, +++ p < 0.0001 versus the AKK-treated group.

Article Snippet: Antibodies against p-p38α MAPK (#4511), HO-1(#43966), NQO-1(#3187) were purchased from Cell Signaling Technology (Danvers, MA, United States).

Techniques: Binding Assay, Biomarker Discovery, Western Blot, Membrane, Expressing

Journal: Frontiers in Microbiology

Article Title: Akkermansia muciniphila alleviates antibiotic- and LPS-induced oxidative stress via the p38α MAPK–Nrf2 signaling axis

doi: 10.3389/fmicb.2026.1753421

Figure Lengend Snippet: A schematic model illustrating the role of the AKK-p38α MAPK-Nrf2 axis and microbial regulation in restoring intestinal homeostasis.

Article Snippet: Antibodies against p-p38α MAPK (#4511), HO-1(#43966), NQO-1(#3187) were purchased from Cell Signaling Technology (Danvers, MA, United States).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Thymoquinone alleviates the accumulation of ROS and pyroptosis and promotes perforator skin flap survival through SIRT1/NF-κB pathway

doi: 10.3389/fphar.2025.1567762

Figure Lengend Snippet: TQ Ameliorates Oxidative Stress and Apoptosis in Rat Perforator Flaps After Ischemia-Reperfusion Injury. (A) Immunohistochemical staining images of SOD1 in the control and TQ groups (scale bar, 50 μm). (B) Quantitative analysis of SOD1 absorbance in the control and TQ groups. (C) Western blotting to detect the expression of oxidative stress-related proteins: eNOS, HO1, and SOD1 in flap tissues of the control and TQ groups. (D) Quantitative analysis of oxidative stress-related protein blots. (E) Immunohistochemical staining images of C-CASP3 in the control and TQ groups (scale bar, 50 μm). (F) Quantitative analysis of C-CASP3 absorbance in the control and TQ groups. (G) Western blotting to detect the expression of apoptosis-related proteins: Bax, C-CASP3, and Bcl-2 in flap tissues of the control and TQ groups. (H) Quantitative analysis of apoptosis-related protein blots. Significance: *p < 0.05 and **p < 0.01. Data are presented as mean ± SEM, with five rats per group.

Article Snippet: Anti-cadherin 5 is provided by Wuhan Boster Biological Technology, Ltd. Primary antibodies against GAPDH, HO1, SOD1, Bax, Bcl-2, GSDMD-N, and VEGF are sourced from The Proteintech Group (Chicago, USA).

Techniques: Immunohistochemical staining, Staining, Control, Western Blot, Expressing

Journal: Frontiers in Pharmacology

Article Title: Thymoquinone alleviates the accumulation of ROS and pyroptosis and promotes perforator skin flap survival through SIRT1/NF-κB pathway

doi: 10.3389/fphar.2025.1567762

Figure Lengend Snippet: EX527 Reverses the Inhibitory Effects of TQ on Pyroptosis, Oxidative Stress, and Apoptosis in Rat Perforator Flaps After Ischemia-Reperfusion Injury. (A) Western blotting to detect the expression of pyroptosis-related proteins: NLRP3, GSDMD-N, Caspase-1, IL-1β, IL-18, and ASC in flap tissues of the control, TQ, and TQ+EX527 groups. (B) Quantitative analysis of pyroptosis-related protein blots. (C) GSDMD-N protein expression levels in flap tissues of rats in the control, TQ, and TQ+EX527 groups measured by immunofluorescence (scale bar: 20 µm). (D) Fluorescence intensity of GSDMD-N expression in each group analyzed using Image J. (E) Western blotting to detect the expression of oxidative stress-related proteins: eNOS, HO1, and SOD1 in flap tissues of the control, TQ, and TQ+EX527 groups. (F) Quantitative analysis of oxidative stress-related protein blots. (G) Western blotting to detect the expression of apoptosis-related proteins: Bax, C-CASP3, and Bcl-2 in flap tissues of the control, TQ, and TQ+EX527 groups. (H) Quantitative analysis of apoptosis-related protein blots. Significance: *p < 0.05 and **p < 0.01. Data are presented as mean ± SEM, with five rats per group.

Article Snippet: Anti-cadherin 5 is provided by Wuhan Boster Biological Technology, Ltd. Primary antibodies against GAPDH, HO1, SOD1, Bax, Bcl-2, GSDMD-N, and VEGF are sourced from The Proteintech Group (Chicago, USA).

Techniques: Western Blot, Expressing, Control, Immunofluorescence, Fluorescence

Journal: Antioxidants

Article Title: Neuron-Derived Sema3B Facilitates Microglial Hematoma Clearance After Intracerebral Hemorrhage

doi: 10.3390/antiox15020220

Figure Lengend Snippet: Sema3B increases HO-1 and TREM2 expression in perihematomal brain tissue and microglia after ICH. ( A – C ) Immunoblotting showing relative HO-1 and TREM2 protein expression in microglia under the different treatments ( n = 4). ( D , E ) Quantitative PCR (qRT-PCR) assessment of HO-1 and TREM2 transcript levels in microglia under the different treatments ( n = 3). ( F – H ) Protein analysis by Western blot for HO-1 and TREM2 in perihematomal tissue from ICH mice on post-ICH day 3 under the different treatments ( n = 4) (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: The antibodies used are as follows: Sema3B (1:2000, R&D Systems, USA), PlexinA1 (1:2000, Abcam, USA), Nrf2 (1:2000, Proteintech, Wuhan, China), TREM2 (1:2000, Proteintech, China), HO-1 (1:2000, Proteintech, China), PI3K (1:2000, Proteintech, China), phospho-PI3K (1:2000, Proteintech, China), AKT (1:2000, Proteintech, China), phospho-AKT (1:2000, Proteintech, China), SYK (1:2000, Proteintech, China), phospho-SYK (1:2000, Proteintech, China), mTOR (1:2000, Proteintech, China), phospho-mTOR (1:2000, Proteintech, China), p65 (1:2000, Proteintech, China), phospho-p65 (1:2000, Proteintech, China), IκB-α (1:2000, Proteintech, China), phospho-IκB-α (1:2000, Proteintech, China), CD206 (1:2000, Proteintech, China), CD86 (1:2000, Abclonal, China), DAP12 (1:2000, CST, Danvers, MA, USA), and GAPDH (1:3000, Proteintech, China).

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

Journal: Antioxidants

Article Title: Neuron-Derived Sema3B Facilitates Microglial Hematoma Clearance After Intracerebral Hemorrhage

doi: 10.3390/antiox15020220

Figure Lengend Snippet: Sema3B upregulates HO-1 and TREM2 expression via Nrf2. ( A , B ) Immunoblotting for relative Nrf2 protein expression in microglia under the different treatments ( n = 3). ( C ) Quantitative PCR (qRT-PCR) assessment of relative Nrf2 transcript levels in microglia under the different treatments ( n = 3). ( D ) Confocal micrographs depicting Nrf2 expression in microglia under the different treatments ( n = 3; scale bar = 10 μm). ( E – G ) Protein analysis by Western blot for HO-1 and TREM2 in microglia under the different treatments ( n = 3) (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: The antibodies used are as follows: Sema3B (1:2000, R&D Systems, USA), PlexinA1 (1:2000, Abcam, USA), Nrf2 (1:2000, Proteintech, Wuhan, China), TREM2 (1:2000, Proteintech, China), HO-1 (1:2000, Proteintech, China), PI3K (1:2000, Proteintech, China), phospho-PI3K (1:2000, Proteintech, China), AKT (1:2000, Proteintech, China), phospho-AKT (1:2000, Proteintech, China), SYK (1:2000, Proteintech, China), phospho-SYK (1:2000, Proteintech, China), mTOR (1:2000, Proteintech, China), phospho-mTOR (1:2000, Proteintech, China), p65 (1:2000, Proteintech, China), phospho-p65 (1:2000, Proteintech, China), IκB-α (1:2000, Proteintech, China), phospho-IκB-α (1:2000, Proteintech, China), CD206 (1:2000, Proteintech, China), CD86 (1:2000, Abclonal, China), DAP12 (1:2000, CST, Danvers, MA, USA), and GAPDH (1:3000, Proteintech, China).

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

Journal: Antioxidants

Article Title: Neuron-Derived Sema3B Facilitates Microglial Hematoma Clearance After Intracerebral Hemorrhage

doi: 10.3390/antiox15020220

Figure Lengend Snippet: Sema3B promotes hematoma absorption after ICH by enhancing PlexinA1-mediated microglial phagocytic function. Exogenous supplementation of Sema3B binds to its receptor PlexinA1, activating the DAP12-dependent signaling pathway (Syk-PI3K-AKT-mTOR) and NRF2 in microglia, thereby increasing the expression of TREM2 and HO-1 to facilitate microglia-mediated hematoma clearance, while suppressing neuroinflammation by inhibiting the NF-κB pathway and reducing pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α). This figure was drawn by Figdraw.

Article Snippet: The antibodies used are as follows: Sema3B (1:2000, R&D Systems, USA), PlexinA1 (1:2000, Abcam, USA), Nrf2 (1:2000, Proteintech, Wuhan, China), TREM2 (1:2000, Proteintech, China), HO-1 (1:2000, Proteintech, China), PI3K (1:2000, Proteintech, China), phospho-PI3K (1:2000, Proteintech, China), AKT (1:2000, Proteintech, China), phospho-AKT (1:2000, Proteintech, China), SYK (1:2000, Proteintech, China), phospho-SYK (1:2000, Proteintech, China), mTOR (1:2000, Proteintech, China), phospho-mTOR (1:2000, Proteintech, China), p65 (1:2000, Proteintech, China), phospho-p65 (1:2000, Proteintech, China), IκB-α (1:2000, Proteintech, China), phospho-IκB-α (1:2000, Proteintech, China), CD206 (1:2000, Proteintech, China), CD86 (1:2000, Abclonal, China), DAP12 (1:2000, CST, Danvers, MA, USA), and GAPDH (1:3000, Proteintech, China).

Techniques: Expressing