anti socs1  (Boster Bio)

Journal: Bioresources and Bioprocessing

Article Title: Propionate-engineered probiotics reduce radiation-induced intestinal damage

doi: 10.1186/s40643-026-01020-9

Figure Lengend Snippet: Engineered probiotic PG mitigated radiation-induced intestinal injury by regulating the SOCS1/JAK2/STAT3 pathway. a Principal Component Analysis (PCA) revealed differences in gene expression levels among groups. b KEGG Enrichment BarPlot for IR vs. IR + PG displayed the number of differential genes in the pathway represented on the horizontal axis, pathway names on the vertical axis, and colors representing KEGG Level 1 classification. c KEGG-enriched bar graph displaying the top 20 KEGG pathways with the smallest P-values. d The results of the KEGG enrichment analysis werw presented as a scatter plot. (e) The heatmap illustrated the relative expression levels of JAK-STAT signaling pathway-related genes in the IR and IR + PG groups. f Western blot analysis demonstrated the protein expression levels of SOCS1, p-JAK2, JAK2, p-STAT3, and STAT3 across groups

Article Snippet: The primary antibodies included anti-SOCS1 (PSH09-68), JAK2 (SY0245), p-JAK2 (SY24-03), STAT3 (SY24-08), p-STAT3 (SZ43-01), and GAPDH (SA30-01), all purchased from HuaBio-Antibodies Co., Ltd (Hangzhou, China).

Techniques: Gene Expression, Expressing, Western Blot

Journal: Science Advances

Article Title: Brain-derived extracellular vesicles circUsp32 polarized macrophages causing acute kidney injury after traumatic brain injury

doi: 10.1126/sciadv.adz1243

Figure Lengend Snippet: ( A ) RNA pull-down assay shows that circUsp32 binds Socs1. ( B ) RIP assay confirms that circUsp32 is enriched by Socs1. IgG served as the negative control. *** P < 0.001. One-way ANOVA followed by Tukey’s multiple comparisons test. ( C ) Predicted three-dimensional structural model indicating potential binding positions for the interaction among circUsp32, Socs1, and IRF7. ( D ) FISH assay reveals colocalization of circUsp32, Socs1, and IRF7. ( E ) Schematic diagram of the Socs1 domain structure. ( F ) RIP results indicate that the Socs1_△SH2 is essential for binding. ( G ) Schematic diagram of circUsp32 mutation sites. ( H ) RNA pull-down assays identify circUsp32 regions required for binding Socs1. ( I and J ) IRF7 expression levels following cotransfection with oe-circ-NC + oe-Socs1-con, oe-circ-NC + oe-Socs1, oe-circUsp32 + oe-Socs1-con, or oe-circUsp32 + oe-Socs1. n = 3 replicates. * P < 0.05; ** P < 0.01. One-way ANOVA followed by Tukey’s multiple comparisons test. ( K and L ) IRF7 expression levels following cotransfection with sh-circ-NC + sh-Socs1-con, sh-circ-NC + sh-Socs1, sh-circUsp32 + sh-Socs1-con, or sh-circUsp32 + sh-Socs1. n = 3 replicates. ** P < 0.01. One-way ANOVA followed by Tukey’s multiple comparisons test. ( M ) 293T cells were cotransfected with the indicated plasmids and treated with MG132 (1 μM, 12 hours). Protein complexes were subject to IP using an anti-Flag antibody and analyzed via immunoblotting with an anti-HA antibody. Total protein levels (input) were assessed via immunoblotting using anti-FLAG, anti-Myc, and anti–β-actin antibodies. All data are presented as means ± SEM.

Article Snippet: The primary antibodies were against the following proteins: TNF-α (1:100; sc-33639; Santa Cruz Biotechnology), IL-6 (1:200; sc-57315; Santa Cruz Biotechnology), IL-1β (1:200; sc-52012; Santa Cruz Biotechnology), Bax (1:1000; 2772S; Cell Signaling Technology), Bcl-2 (1:1000; 2764S; Cell Signaling Technology), cleaved caspase-3 (1:1000; 9661S; Cell Signaling Technology), Socs1 (1:100; sc-518028; Santa Cruz Biotechnology), IRF7 (1:1000; ET1610-89; HUABIO), Oat1 (1:500; 26574-1-AP; Proteintech), Occludin (1:2000; 27260-1-AP; Proteintech), Claudin-1 (1:1000; ab307692; Abcam), and β-actin (1:20,000; 66009-1-Ig; Proteintech).

Techniques: Pull Down Assay, Negative Control, Binding Assay, Mutagenesis, Expressing, Cotransfection, Western Blot

Journal: Science Advances

Article Title: Brain-derived extracellular vesicles circUsp32 polarized macrophages causing acute kidney injury after traumatic brain injury

doi: 10.1126/sciadv.adz1243

Figure Lengend Snippet: ( A to C ) Socs1 expression levels were detected via WB in Control, TBI-EVs, TBI-EVs + oe-circ-NC, or TBI-EVs + sh-circ-NC, and TBI-EVs + oe-circUsp32 or TBI-EVs + sh-circUsp32 groups. n = 3. *** P < 0.001, n.s., not significant. One-way ANOVA followed by Tukey’s multiple comparisons test. ( D ) Representative double IF images show colocalization of CD68, Socs1, IRF7, iNOS, and CD206 in cells after overexpression or knockdown of Socs1. ( E ) Flow cytometry quantification of pro-inflammatory (iNOS+) and anti-inflammatory (CD206+) macrophages following oe-Socs1 or sh-Socs1 transfection in RAW 264.7 cells. ( F to H ) WB analysis showing IRF7 expression levels in cells transduced oe-circUsp32/sh-circUsp32 alone or cotransduced with oe-Socs1/sh-Socs1 for 48 hours, followed by TBI-EV treatment for 3 hours. n = 3. * P < 0.05; ** P < 0.01; **** P < 0.0001. One-way ANOVA followed by Tukey’s multiple comparisons test. ( I ) Heatmap shows mRNA expression levels of pro-inflammatory ( Tnf- α, Il-6 , and Il-1 β) and anti-inflammatory ( Arg-1 and Il-10 ) macrophage markers via qRT-PCR after transduction oe-circUsp32 alone or cotransduced with oe-Socs1 for 48 hours, followed by TBI-EV treatment for 3 hours. ( J ) Heatmap shows mRNA expression levels of pro-inflammatory ( Tnf- α, Il-6 , and Il-1 β) and anti-inflammatory ( Arg-1 and Il-10 ) macrophage markers via qRT-PCR after transduction sh-circUsp32 alone or cotransduced with sh-Socs1 for 48 hours, followed by TBI-EV treatment for 3 hours. All data are presented as means ± SEM.

Article Snippet: The primary antibodies were against the following proteins: TNF-α (1:100; sc-33639; Santa Cruz Biotechnology), IL-6 (1:200; sc-57315; Santa Cruz Biotechnology), IL-1β (1:200; sc-52012; Santa Cruz Biotechnology), Bax (1:1000; 2772S; Cell Signaling Technology), Bcl-2 (1:1000; 2764S; Cell Signaling Technology), cleaved caspase-3 (1:1000; 9661S; Cell Signaling Technology), Socs1 (1:100; sc-518028; Santa Cruz Biotechnology), IRF7 (1:1000; ET1610-89; HUABIO), Oat1 (1:500; 26574-1-AP; Proteintech), Occludin (1:2000; 27260-1-AP; Proteintech), Claudin-1 (1:1000; ab307692; Abcam), and β-actin (1:20,000; 66009-1-Ig; Proteintech).

Techniques: Expressing, Control, Over Expression, Knockdown, Flow Cytometry, Transfection, Quantitative RT-PCR, Transduction

Journal: Science Advances

Article Title: Brain-derived extracellular vesicles circUsp32 polarized macrophages causing acute kidney injury after traumatic brain injury

doi: 10.1126/sciadv.adz1243

Figure Lengend Snippet: ( A ) Serum cystatin, ( B ) BUN, and ( C ) Scr levels were detected by ELISA in Sham, TBI, TBI-EVs, and TBI-EVs sh-circUsp32 mice. n = 5 mice per group. * P < 0.05; ** P < 0.01; **** P < 0.0001; n.s., not significant. One-way ANOVA followed by Tukey’s multiple comparisons test. ( D ) Urinary NGAL and ( E ) KIM-1 levels were detected via ELISA in Sham, TBI, TBI-EVs, and TBI-EVs sh-circUsp32 mice. n = 5 mice per group. * P < 0.05; ** P < 0.01; **** P < 0.0001; n.s., not significant. One-way ANOVA followed by Tukey’s multiple comparisons test. ( F ) Representative H&E, PAS, and TUNEL staining images of renal structures in Sham, TBI, TBI-EVs, and TBI-EVs sh-circUsp32 mice. ( G ) Representative double IF images display the colocalization of Kim-1, IRF7, Socs1, iNOS, and CD206 in mice kidney following TBI-EVs or TBI-EVs sh-circUsp32 injection. ( H ) Flow cytometry quantification of pro-inflammatory (iNOS+) and anti-inflammatory (CD206+) macrophages in renal immune cell suspensions in Sham, TBI, TBI-EVs, and TBI-EVs sh-circUsp32 mice. ( I and J ) WB analysis of renal expression levels of IRF7, Socs1, TNF-α, IL-6, IL-1β, Bax, Bcl-2, and cleaved caspase-3 in Sham, TBI, TBI-EVs, and TBI-EVs sh-circUsp32 mice. n = 5 mice per group. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; n.s., not significant. One-way ANOVA followed by Tukey’s multiple comparisons test. All data are presented as means ± SEM.

Article Snippet: The primary antibodies were against the following proteins: TNF-α (1:100; sc-33639; Santa Cruz Biotechnology), IL-6 (1:200; sc-57315; Santa Cruz Biotechnology), IL-1β (1:200; sc-52012; Santa Cruz Biotechnology), Bax (1:1000; 2772S; Cell Signaling Technology), Bcl-2 (1:1000; 2764S; Cell Signaling Technology), cleaved caspase-3 (1:1000; 9661S; Cell Signaling Technology), Socs1 (1:100; sc-518028; Santa Cruz Biotechnology), IRF7 (1:1000; ET1610-89; HUABIO), Oat1 (1:500; 26574-1-AP; Proteintech), Occludin (1:2000; 27260-1-AP; Proteintech), Claudin-1 (1:1000; ab307692; Abcam), and β-actin (1:20,000; 66009-1-Ig; Proteintech).

Techniques: Enzyme-linked Immunosorbent Assay, TUNEL Assay, Staining, Injection, Flow Cytometry, Expressing

Journal: Science Advances

Article Title: Brain-derived extracellular vesicles circUsp32 polarized macrophages causing acute kidney injury after traumatic brain injury

doi: 10.1126/sciadv.adz1243

Figure Lengend Snippet: CircUsp32 acts as a molecular sponge for Socs1, leading to up-regulation of IRF7 subsequent pro-inflammatory macrophage polarization, which contributes to AKI following TBI. Injured renal tubular epithelial cells may down-regulate Oat1 expression, resulting in elevated levels in peripheral blood that reach the central nervous system and trigger neuroinflammation.

Article Snippet: The primary antibodies were against the following proteins: TNF-α (1:100; sc-33639; Santa Cruz Biotechnology), IL-6 (1:200; sc-57315; Santa Cruz Biotechnology), IL-1β (1:200; sc-52012; Santa Cruz Biotechnology), Bax (1:1000; 2772S; Cell Signaling Technology), Bcl-2 (1:1000; 2764S; Cell Signaling Technology), cleaved caspase-3 (1:1000; 9661S; Cell Signaling Technology), Socs1 (1:100; sc-518028; Santa Cruz Biotechnology), IRF7 (1:1000; ET1610-89; HUABIO), Oat1 (1:500; 26574-1-AP; Proteintech), Occludin (1:2000; 27260-1-AP; Proteintech), Claudin-1 (1:1000; ab307692; Abcam), and β-actin (1:20,000; 66009-1-Ig; Proteintech).

Techniques: Expressing