csb el026133mo  (Cusabio)

Journal: Cell Reports Medicine

Article Title: A combined “eat me/don’t eat me” strategy based on exosome for acute liver injury treatment

doi: 10.1016/j.xcrm.2025.102033

Figure Lengend Snippet: Efficacy of Wnt2 mRNA-loaded exosome in vitro (A) Wnt2 mRNA expression in Exo and Wnt2@Exo CD47 detected by RT-qPCR. n = 3. (B) CLSM images showing the internalization of DiO-labeled Exo and Wnt2@Exo CD47 by HUVEC (left), 293T (meddle), and primary LSEC (right). Red: TRITC-phalloidin-labeled F-actin. Green: DiO-labeled Exo and Wnt2@Exo CD47 . Blue: DAPI-labeled nuclei. Scale bar: 20 μm (C) ELISA analysis of Wnt2 concentration in culture supernatant of HUVEC (left), 293T (meddle), and primary LSEC (right) at 48 h after incubation with Exo and Wnt2@Exo CD47 . n = 3. (D) RT-qPCR shows higher Wnt2 mRNA transcript levels after in vitro delivery of Wnt2 mRNA from Wnt2@Exo CD47 (Exo vs. Wnt2@Exo CD47 ) in 48 h. n = 3. (E) Following incubation with conditioned medium for 24 h, RT-qPCR was employed to analyze the changes in downstream target genes of the Wnt pathway in the primary hepatocytes. n = 3. Data are represented as mean ± SD (error bars) from biological replicates. Statistical analyses, n.s., not significant, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Statistical significance was determined by unpaired Student’s t test.

Article Snippet: Mouse Protein Wnt-2 (WNT2) ELISA kit , CUSABIO , CSB-EL026133MO.

Techniques: In Vitro, Expressing, Quantitative RT-PCR, Labeling, Enzyme-linked Immunosorbent Assay, Concentration Assay, Incubation

Journal: Cell Reports Medicine

Article Title: A combined “eat me/don’t eat me” strategy based on exosome for acute liver injury treatment

doi: 10.1016/j.xcrm.2025.102033

Figure Lengend Snippet: The engineered exosome treatment alleviates DMN/APAP-induced acute liver injury (A) Schematic illustration of the experimental procedure. (B) The overall appearance of the livers in DMN (up)- and APAP (down)-induced acute liver injury mice after tail vein administration of PBS, Exo, or RLTR-Wnt2@Exo CD47 . (C) Serum ALT, AST, ALB, and TBIL of mice with DMN-induced liver injury at 48 h after PBS, Exo, or RLTR-Wnt2@Exo CD47 administration. n = 6. ALB, albumin; ALT, alanine aminotransferase; AST, aspartate aminotransferase; TBIL, total bilirubin. (D) Serum ALT, AST, ALB, and TBIL of mice with APAP-induced liver injury at 48 h after PBS, Exo, or RLTR-Wnt2@Exo CD47 administration. n = 6. ALB, albumin; ALT, alanine aminotransferase; AST, aspartate aminotransferase; TBIL, total bilirubin. (E and F) Ki67 immunofluorescence staining of DMN (E)- and APAP (F)-injured liver after PBS, Exo, or RLTR-Wnt2@Exo CD47 administration. Red: ki67. Blue: nuclei. Scale bar: 100 μm. (G) H&E staining of liver collected from DMN (up) and APAP (down) mice after treated with PBS, Exo, or RLTR-Wnt2@Exo CD47 . Scale bar: 200 μm. (H) Quantitative analysis of the Ki67 + hepatocytes ratio. n = 6. (I) Quantitative analysis of the necrotic areas. n = 6. (J) The heatmap based on the differently expressed genes between livers from control and RLTR-Wnt2@Exo CD47 group. Data were obtained from biological replicates. CT: control group, R: RLTR-Wnt2@Exo CD47 group. (K) GSEA revealing the enrichment of differently expressed genes in the Wnt, positive regulation of cell-cycle process, liver regeneration, and hepatocyte differentiation signaling pathways. n = 3. CT, control group; R, RLTR-Wnt2@Exo CD47 group. Data are represented as mean ± SD (error bars) from biological replicates. Statistical analyses, n.s., not significant, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Statistical significance was determined by one-way ANOVA. See also Figures S3–S6 .

Article Snippet: Mouse Protein Wnt-2 (WNT2) ELISA kit , CUSABIO , CSB-EL026133MO.

Techniques: Immunofluorescence, Staining, Control, Protein-Protein interactions

Journal: Cell Reports Medicine

Article Title: A combined “eat me/don’t eat me” strategy based on exosome for acute liver injury treatment

doi: 10.1016/j.xcrm.2025.102033

Figure Lengend Snippet: Safety evaluation of the engineered exosome in vitro and in vivo (A) Cell viability of bEnd.3 was analyzed by CCK8 assay after incubation with different concentrations of Exo or RLTR-Wnt2@Exo CD47 at 12, 24, and 48 h. n = 3. (B) Scheme showing the treatment timeline of PBS, Exo, and RLTR-Wnt2@Exo CD47 . (C–E) Quantitative analysis of serological detection indicators about liver function (C), renal function (D), and myocardial enzymes (E) in serum of mice with different treatment. PBS: n = 4, Exo: n = 5, RLTR-Wnt2@Exo CD47 : n = 4. ALB, albumin; ALT, alanine aminotransferase; AST, aspartate aminotransferase; TBIL, total bilirubin; BUN, blood urea nitrogen; CREA, creatine; LDH, lactate dehydrogenase; LDH1, lactate dehydrogenase 1. (F) H&E-stained images of the major organs (heart, liver, spleen, lung, and kidney) collected from mice in different treatment groups after sacrifice. Scale bar: 100 μm. (G) Schematic showed the working model of this study. Exosomes are isolated from bEnd.3 culture medium and engineered with a dual-targeting “eat me/don’t eat me” strategy, enabling them to evade macrophage phagocytosis while specifically targeting LSEC. Engineered exosomes deliver Wnt2 mRNA to LSEC, enhancing the release of Wnt2 into the sinusoidal niche. This activates the Wnt/β-catenin signaling pathway in hepatocytes, driving liver regeneration following acute liver injury. Data are represented as mean ± SD (error bars) from biological replicates. Statistical analyses, n.s., not significant, ∗ p < 0.05, ∗∗ p < 0.01. Statistical significance was determined by one-way ANOVA.

Article Snippet: Mouse Protein Wnt-2 (WNT2) ELISA kit , CUSABIO , CSB-EL026133MO.

Techniques: In Vitro, In Vivo, CCK-8 Assay, Incubation, Staining, Isolation

Journal: Cell Reports Medicine

Article Title: A combined “eat me/don’t eat me” strategy based on exosome for acute liver injury treatment

doi: 10.1016/j.xcrm.2025.102033

Figure Lengend Snippet:

Article Snippet: Mouse Protein Wnt-2 (WNT2) ELISA kit , CUSABIO , CSB-EL026133MO.

Techniques: Virus, Plasmid Preparation, Recombinant, Sterility, Modification, CCK-8 Assay, SYBR Green Assay, Enzyme-linked Immunosorbent Assay, TUNEL Assay, Apoptosis Assay, Software

Journal: NPJ Regenerative Medicine

Article Title: IRES-mediated Wnt2 translation in apoptotic neurons triggers astrocyte dedifferentiation

doi: 10.1038/s41536-022-00248-1

Figure Lengend Snippet: a Double-immunostaining of NeuN/Wnt2 in normal cortex. Bar = 50 μm. b , c Western-blotting of Wnt2 at different time points in ischemic cortex. Notice that Wnt2 protein increases quickly after ischemia. * P = 0.026 (1 h), 0.028 (2 h), 0.024 (4 h), 0.048 (8 h), 0.043 (12 h). One-way ANOVA followed by Bonferroni’s post hoc comparisons tests. N = 4 mice per group. d Combination of TUNEL staining and Wnt2 immunostaining at 24 hpi. Wnt2 high immunoreactivity overlapped well with TUNEL-staining (arrowheads). Dashed line indicates lesion border and asterisk indicates lesion area. Bar = 30 μm. e Representative image of TUNEL/Wnt2/NeuN-triple immunostaining in the lesion area. Bar = 10 μm. f , g Western-blotting of Wnt2 in conditioned medium of normal cultured (NNCM), apoptotic (ANCM), Wnt2 shRNA pretreated apoptotic (shRNA ANCM) wild type and caspase-3 −/− neurons. Apoptotic neurons increase Wnt2 release into the culture medium, and this increase is compromised by caspase-3 ablation and Wnt2 shRNA. BSA was used as internal control. * P NNCM vs ANCM = 0.012, * P ANCM vs ShRNA-ANCM = 0.014. One-way ANOVA followed by Bonferroni’s post hoc comparisons tests. N = 3 batches of cells. h Western-blotting of active β-catenin in the cytoplasmic and nuclear protein of astrocytes treated by NNCM, ANCM, AICM or shRNA ANCM. ANCM greatly increases the level of β-catenin in nuclear protein. LMNB1 was used as nuclear protein control. i Double-immunostaining of GFAP/β-gal in Topgal mice pretreated with Wnt2 shRNA. Arrows point to representative double-positive cells. Bar = 50 μm. j Western-blotting of β-gal and Axin2 in ischemic cortex of Topgal mice treated with Wnt2 shRNA or control. Left: * P ipsi vs ipsi+Wnt2-shRNA = 0.038, * P ipsi+Con-shRNA vs ipsi+Wnt2-shRNA = 0.041, Right: * P ipsi vs ipsi+Wnt2-shRNA = 0.013, * P ipsi+Con-shRNA vs ipsi+Wnt2-shRNA = 0.016. One-way ANOVA followed by Bonferroni’s post hoc comparisons tests. N = 3 mice per group. Notice the less induction of β-gal and Axin2 in Wnt2 shRNA pretreated cortex. Mean ± standard error (SE). Asterisks indicate the comparison with control group. Asterisks with bars connecting two groups indicate difference between these two groups.

Article Snippet: The concentrations of Wnt2 in the conditioned medium of control neurons, neurons treated with OGD, and neurons treated with OGD and Wnt2 shRNA were determined using an ELISA kit (Cusabio, CSB-EL026133MO) according to the manufacturer’s instruction.

Techniques: Double Immunostaining, Western Blot, TUNEL Assay, Staining, Immunostaining, Triple Immunostaining, Cell Culture, shRNA, Control, Comparison

Journal: NPJ Regenerative Medicine

Article Title: IRES-mediated Wnt2 translation in apoptotic neurons triggers astrocyte dedifferentiation

doi: 10.1038/s41536-022-00248-1

Figure Lengend Snippet: a Representative images of Nestin/GFAP double-immunostaining in the bilateral cortice with two ischemic injures infected by lentivirus expressing control shRNA (right) and Wnt2 shRNA (left), respectively. Bars = 200 μm in upper panel. Bar = 100 μm in enlarged view. b Western-blotting of Nestin and Sox2 in Wnt2 shRNA treated mice at 7 dpi. Wnt2 shRNA dramatically suppressed the expression of Nestin and Sox2 without obviously affecting the expression of GFAP. N = 5 mice per group. c Double-immunostaining and quantification of DCX/BrdU in ischemic cortices treated by control shRNA or Wnt2 shRNA. Arrowheads point to double-positive cells. Arrows point to magnified cells. Bar = 20 μm. ** P = 0.0087. Two-tailed Student’s t test. N = 4 mice per group. d – f Forelimb activity assay of mice treated with control shRNA or Wnt2 shRNA. From 2 weeks on, the sliding scores and the asymmetric index of forelimb activity in Wnt2 shRNA treated group were much higher than control group, indicating worse functional recovery. N = 5 mice per group. P < 0.0001 (2w, 3w, 4w and 5w). Two-way RM ANOVA. a – c , asterisks with bars connecting two groups indicate difference between these two groups. e , f , P values indicate comparison of experimental group with control group of same time point. Mean ± standard error (SE).

Article Snippet: The concentrations of Wnt2 in the conditioned medium of control neurons, neurons treated with OGD, and neurons treated with OGD and Wnt2 shRNA were determined using an ELISA kit (Cusabio, CSB-EL026133MO) according to the manufacturer’s instruction.

Techniques: Double Immunostaining, Infection, Expressing, Control, shRNA, Western Blot, Two Tailed Test, Activity Assay, Functional Assay, Comparison

Journal: NPJ Regenerative Medicine

Article Title: IRES-mediated Wnt2 translation in apoptotic neurons triggers astrocyte dedifferentiation

doi: 10.1038/s41536-022-00248-1

Figure Lengend Snippet: a Western-blotting of Wnt2 in ischemic cortex infected by pLenti-Luci (control) and Wnt2. b Western-blotting of Nestin and double-immunostaining of Nestin/Sox2 in ischemic cortex infected by pLenti-Luci (control) and Wnt2. Bar = 50 μm. N = 4 mice per group. c Representative images of Nestin/GFAP double-immunostaining in the bilateral cortices at 14 dpi with two ischemic injures infected by pLenti-Luci (left) and Wnt2 (right) respectively. Bar = 200 μm in upper panel. Bar = 50 μm in enlarged view. N = 4 mice per group. d Double-immunostaining and quantification of DCX/BrdU in ischemic cortex infected by pLenti-Luci and Wnt2. N = 3 mice per group. ** P = 0.0075. Two-tailed Student’s t test. Bar = 50 μm. e Double-immunostaining and quantification of NeuN/BrdU in ischemic cortices infected by lentivirus expressing luciferase (control) or Wnt2. S1 and S2 are two representative sections showing one NeuN/BrdU-positive cell in each section. Arrows point to double-stained cells. Bar = 30 μm, and 5 μm in insert. *** P < 0.001. Two-tailed Student’s t test. N = 4 mice per group. Mean ± standard error (SE). Asterisks with bars connecting two groups indicate difference between these two groups.

Article Snippet: The concentrations of Wnt2 in the conditioned medium of control neurons, neurons treated with OGD, and neurons treated with OGD and Wnt2 shRNA were determined using an ELISA kit (Cusabio, CSB-EL026133MO) according to the manufacturer’s instruction.

Techniques: Western Blot, Infection, Control, Double Immunostaining, Two Tailed Test, Expressing, Luciferase, Staining

Journal: NPJ Regenerative Medicine

Article Title: IRES-mediated Wnt2 translation in apoptotic neurons triggers astrocyte dedifferentiation

doi: 10.1038/s41536-022-00248-1

Figure Lengend Snippet: a Combination of TUNEL staining and Wnt2 immunostaining (left panels), and Western-blotting of Wnt2 in the ischemic cortex of WT and caspase-3 −/− mice (right panels). Expression of Wnt2 in the injured WT cortex is significantly increased, but remains unchanged in caspase-3 −/− cortex. Dashed lines indicate lesion border. Bar = 30 μm. ** P = 0.0061 (left panel), ** P = 0.0019 (right panel). One-way ANOVA followed by Bonferroni’s post hoc comparisons tests. N = 3 mice per group. b Representative images of Nestin/β-gal double immunostaining (left panels), and Western-blotting of β-gal in the ischemic cortex of Topgal mice with or without caspase-3 mutation at 5 dpi (right panels). The expression of β-gal in the ischemic cortex of Topgal:caspase-3 +/+ mice is greatly enhanced, whereas it shows no notable change in Topgal:caspase-3 −/− mice. Bar = 30 μm. ** P = 0.0096 (left panel), ** P = 0.0015 (right panel). One-way ANOVA followed by Bonferroni’s post hoc comparisons tests. N = 3–4 mice per group. c Double staining of BrdU with Nestin, and Western-blotting of Nestin in the ischemic WT and caspase-3 −/− cortex. Notice the lower level of Nestin in the ischemic caspase-3 −/− cortex. ** P = 0.001 (left panel), ** P = 0.0012 (right panel). One-way ANOVA followed by Bonferroni’s post hoc comparisons tests. N = 3–4 mice per group. d Double staining and quantification of BrdU/DCX. The numbers of BrdU/DCX-positive cells is significantly decreased in the caspase-3 −/− cortex. Bars = 30 μm. ** P = 0.0072 (left panel), ** P = 0.0021 (right panel). Two-tailed Student’s t test. N = 3–4 per group. Arrows in ( b , d ) point to double-positive cells and arrowheads in ( d ) point to magnified double-positive cells. Mean ± standard error (SE). Asterisks with bars connecting two groups indicate difference between these two groups.

Article Snippet: The concentrations of Wnt2 in the conditioned medium of control neurons, neurons treated with OGD, and neurons treated with OGD and Wnt2 shRNA were determined using an ELISA kit (Cusabio, CSB-EL026133MO) according to the manufacturer’s instruction.

Techniques: TUNEL Assay, Staining, Immunostaining, Western Blot, Expressing, Double Immunostaining, Mutagenesis, Double Staining, Two Tailed Test

Journal: NPJ Regenerative Medicine

Article Title: IRES-mediated Wnt2 translation in apoptotic neurons triggers astrocyte dedifferentiation

doi: 10.1038/s41536-022-00248-1

Figure Lengend Snippet: a , b Double-immunostaining of NeuN/Wnt2, Wnt2/CC3 in control and injured cortex of Macaca Mulatta at 24 hpi. Bar = 30 μm. c , d Double-immunostaining of NeuN/Wnt2, Wnt2/CC3 in human cortical autopsy tissue (control and 24 hpi). Bar = 30 μm. Notice the higher Wnt2-immunoreactivity in CC3-positive cells in both Macaca Mulatta and human cortex. Arrows point to representative double-positive cells. *** P < 0.001. Two-tailed Student’s t test. N = 3 samples from 2 monkeys or patients per group. Mean ± standard error (SE). Asterisks with bars connecting two groups indicate difference between these two groups.

Article Snippet: The concentrations of Wnt2 in the conditioned medium of control neurons, neurons treated with OGD, and neurons treated with OGD and Wnt2 shRNA were determined using an ELISA kit (Cusabio, CSB-EL026133MO) according to the manufacturer’s instruction.

Techniques: Double Immunostaining, Control, Two Tailed Test

Journal: NPJ Regenerative Medicine

Article Title: IRES-mediated Wnt2 translation in apoptotic neurons triggers astrocyte dedifferentiation

doi: 10.1038/s41536-022-00248-1

Figure Lengend Snippet: a Real time RT-PCR of Wnt2 in OGD treated neurons. b Western-blotting of Wnt2 in apoptotic neurons treated with PD169316. N = 3 batches of cells. c Western-blotting and quantification of Wnt2 in apoptotic neurons treated with CHX. Notice that OGD does not induce Wnt2 mRNA transcription and that CHX blocks OGD-induced Wnt2 protein up-regulation. * P con vs OGD = 0.011, * P OGD VS OGD+CHX = 0.031. One-way ANOVA followed by Bonferroni’s post hoc comparisons tests. N = 3 batches of cells. d Sequence alignment of mouse, Macaca Mulatta and human Wnt2-5UTR. e Secondary structure of mouse Wnt2-5UTR. Blue frames in ( d ) and ( e ) show the largest stem-loop and the corresponding sequence. The sequence underlined by black dots (in d ) was replaced by the sequence below highlighted with yellow color to make Wnt2-5UTR mutant construct in ( f ). f , g Bicistronic luciferase reporter assay of XIAP-5UTR, Wnt2-5UTR, and Wnt2-5UTR mutant under control and OGD condition. ** P XIAP = 0.0022, ** P Wnt2 = 0.0019. One-way ANOVA followed by Bonferroni’s post hoc comparisons tests. N = 3 batches of cells. Notice that Wnt2-5UTR initiates OGD-induced Wnt2 translation and mutation at the largest loop abolishes this translation. Mean ± standard error (SE). Asterisks with bars connecting two groups indicate difference between these two groups.

Article Snippet: The concentrations of Wnt2 in the conditioned medium of control neurons, neurons treated with OGD, and neurons treated with OGD and Wnt2 shRNA were determined using an ELISA kit (Cusabio, CSB-EL026133MO) according to the manufacturer’s instruction.

Techniques: Quantitative RT-PCR, Western Blot, Sequencing, Mutagenesis, Construct, Luciferase, Reporter Assay, Control

Journal: NPJ Regenerative Medicine

Article Title: IRES-mediated Wnt2 translation in apoptotic neurons triggers astrocyte dedifferentiation

doi: 10.1038/s41536-022-00248-1

Figure Lengend Snippet: a RIP assay. Notice that DAP5 could bind to the Wnt2 5UTR under OGD. *** P < 0.001. One-way ANOVA. N = 3 batches of cells. b IRES activity of Wnt2 5UTR with or without DAP5 shRNA under OGD. DAP5 shRNA effectively blocked the OGD-induced IRES activity. ** P = 0.0052. One-way ANOVA followed by Bonferroni’s post hoc comparisons tests. N = 3 batches of cells. c Western-blotting of Wnt2 in the contralateral and ipsilateral cortex infected by lentivirus expressing DAP5 shRNA or control shRNA. * P = 0.039. One-way ANOVA followed by Bonferroni’s post hoc comparisons tests. N = 3 mice per group. d Double-immunostaining and quantification of GFAP/β-gal in ischemic cortex treated by control shRNA and DAP5 shRNA. Notice the attenuated expression of β-gal in DAP5 shRNA treated cortex. ** P = 0.0081. Two-tailed Student’s t test. N = 3 mice per group. e Double-immunostaining and quantification of GFAP/Nestin in ischemic cortex treated by control shRNA and DAP5 shRNA. Ischemia-induced Nestin was reduced in DAP5 shRNA treated cortex. ** P = 0.0097. Two-tailed Student’s t test. N = 3 mice per group. f Double-immunostaining of DCX/BrdU in ischemic cortex treated by control shRNA and DAP5 shRNA. Notice the decreased number of DCX/BrdU-positive cells in DAP5 shRNA treated cortex. Arrowheads point to double-positive cells. Arrows points to magnified cells. Bar = 30 μm. ** P = 0.0063. Two-tailed Student’s t test. N = 3 mice per group. Mean ± standard error (SE). Asterisks with bars connecting two groups indicate difference between these two groups.

Article Snippet: The concentrations of Wnt2 in the conditioned medium of control neurons, neurons treated with OGD, and neurons treated with OGD and Wnt2 shRNA were determined using an ELISA kit (Cusabio, CSB-EL026133MO) according to the manufacturer’s instruction.

Techniques: Activity Assay, shRNA, Western Blot, Infection, Expressing, Control, Double Immunostaining, Two Tailed Test

Journal: NPJ Regenerative Medicine

Article Title: IRES-mediated Wnt2 translation in apoptotic neurons triggers astrocyte dedifferentiation

doi: 10.1038/s41536-022-00248-1

Figure Lengend Snippet: Following cerebral ischemia, apoptotic neurons up-regulate Wnt2 protein via IRES mediated protein translation. Released Wnt2 activates Wnt/β-catenin signaling in reactive astrocyte and triggers astrocytic dedifferentiation, which facilitated cortical neurogenesis.

Article Snippet: The concentrations of Wnt2 in the conditioned medium of control neurons, neurons treated with OGD, and neurons treated with OGD and Wnt2 shRNA were determined using an ELISA kit (Cusabio, CSB-EL026133MO) according to the manufacturer’s instruction.

Techniques: