Journal: Frontiers in Molecular Neuroscience

Article Title: Artemin and an Artemin-Derived Peptide, Artefin, Induce Neuronal Survival, and Differentiation Through Ret and NCAM

doi: 10.3389/fnmol.2019.00047

Figure Lengend Snippet: Inhibition of ARTN-induced (A) and artefin-induced (B) neurite outgrowth in CGNs by anti-RET antibody. CGNs were pre-treated directly after plating for 1 h with anti-RET antibody or control IgG in concentrations 4.1 μg/ml before stimulating with ARTN (0.042 nM) or artefin (0.47 μM). Student’s paired t -test was used for statistical evaluation of the results. + Indicates p -values for comparison of the neuritogenic effect of ARTN or artefin to the untreated control set at 100%, while ∗ shows p -values for the comparison of the inhibition effect of RET antibody to the control IgG. + p < 0.05; ++ p < 0.01; ∗ p < 0.05; ∗∗ p < 0.01.

Article Snippet: For the RET inhibition assay, the inhibitory goat anti-mouse RET antibody (4.1 μg/ml; R&D systems, Abingdon, United Kingdom) or control goat IgG (4.1 μg/ml; Santa Cruz Biotechnology, Dallas, TX, United States) were added to neurons 1 h before stimulation with ARTN (0.042 nM) or artefin (4.2 μM).

Techniques: Inhibition, Control, Comparison

Journal: Cell Death Discovery

Article Title: TRIM27-controlled endothelium-derived exosomes play a central role in podocyte injury in diabetic kidney disease

doi: 10.1038/s41420-026-02953-y

Figure Lengend Snippet: A Double immunofluorescence staining of TRIM27 protein in glomerular endothelial cells of DKD patients. Kidney sections were costained for TRIM27 (green) and specific endothelial cell marker CD31 (red). Scale bars: 10 μm. B Significant positive correlation was observed between TRIM27 expression and proteinuria in DKD patients ( n = 36 DKD patients). C , D The ELISA assays showed serum contents of syndecan-1 and VCAM-1 increased in DKD patients. *** P < 0.001 vs. Normal group ( n = 24 normal controls and 36 DKD patients). E , F Significant positive correlation was observed between TRIM27 expression and syndecan-1 and VCAM-1 serum levels in DKD patients ( n = 36 DKD patients). G Expression of TRIM27 (green) in glomerular endothelial cells (specific marker, CD31; red) of STZ-induced diabetec mice detected by immunofluorescence. Scale bars: 10 μm. H Expression of syndecan-1 in the glomerulus of STZ mice detected by immunofluorescence. Scale bars: 25 μm. I Significant negative correlation was observed between TRIM27 and syndecan-1 expression in STZ mice ( n = 6 mice, 10 glomeruli per mouse). J – L Western blot assay showed TRIM27 expression increased in HRGECs treated with HG or TGF-β1. ** P < 0.01, **** P < 0.0001 vs. 0 h group ( n = 3). M IF assay showed that the expression of TRIM27 increased in HRGECs treated with TGF-β1. Scale bars: 25 μm. N – P Western blot assay showed VCAM-1 expression increased in HRGECs treated with HG or TGF-β1. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 vs. 0 h group ( n = 3). Q , R The ELISA assays showed the contents of syndecan-1 and VCAM-1 increased in cultures supernatant of HRGECs treated with HG for 24 h. ** P < 0.01, **** P < 0.0001 vs. 0 h group ( n = 3). S NO level increased in the HRGECs after treated with HG. ** P < 0.01, **** P < 0.0001 vs. 0 h group ( n = 3). Student’s t test and Bonferroni’s correction were performed to analyze statistical significance. Pearson’s method was used for correlation analysis. Data are the mean ± SEM.

Article Snippet: Then, the membranes were incubated overnight at 4 °C with 1:1000 dilutions of antibodies against TRIM27 (Proteintech, #12205-1-AP), VCAM-1 (Abcam, #ab134047, RRID: AB_2721053), Janus kinase 2 (JAK2; Proteintech, #17670-1-AP, RRID: AB_2811138), p-JAK2 (Y1007) (Abcam, #ab195055), signal transducer and activator of transcription 3 (STAT3; Proteintech, #60199-1-Ig, RRID: AB_10913811), p-STAT3 (Tyr705) (CST, #9145, RRID: AB_2491009), nephrin (Abcam, #ab58968, RRID: AB_944400), podocin (Abcam, #ab50339, RRID: AB_882097), CD63 (Santa Cruz Biotechnology, Dallas, TX, USA, #sc-5275, RRID: AB_627877), CD9 (Santa Cruz Biotechnology, #sc-13118, RRID: AB_627213), TSG101 (Proteintech, #28283-1-AP, RRID: AB_2881104), Calnexin (Proteintech, #10427-2-AP, RRID: AB_2069033), Rab27a (Proteintech, #17817-1-AP, RRID: AB_2176728), PTEN (Abcam, #ab32199, RRID: AB_777535), Akt (Proteintech, #60203-2-Ig, RRID: AB_10912803), and p-Akt (S473) (Abcam, #ab81283, RRID: AB_2224551).

Techniques: Double Immunofluorescence Staining, Marker, Expressing, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Western Blot

Journal: Cell Death Discovery

Article Title: TRIM27-controlled endothelium-derived exosomes play a central role in podocyte injury in diabetic kidney disease

doi: 10.1038/s41420-026-02953-y

Figure Lengend Snippet: A – F Western blot assay showed TRIM27 and VCAM-1 expression decreased in HRGECs treated with shTRIM27 and HG or TGF-β1 for 24 h. * P < 0.05, *** P < 0.001, **** P < 0.0001 vs. control group, # P < 0.05, #### P < 0.0001 vs. HG+shNC group, ## P < 0.01, ### P < 0.001 vs. TGF-β1+shNC group ( n = 3). G , H ELISA assays showed contents of syndecan-1 and VCAM-1 decreased in culture supernatants after TRIM27 knockdown in HRGECs and treatment with HG for 24 h. * P < 0.05, **** P < 0.0001 vs. control group, ## P < 0.01, #### P < 0.0001 vs. HG+shNC group ( n = 3). I – K IF assay showed that downregulation of TRIM27 increased VE-cadherin, ZO-1, and syndecan-1 expression in HRGECs treated with TGF-β1 for 24 h. Scale bars: 25 μm. Bonferroni’s correction was performed to analyze statistical significance. Data are the mean ± SEM.

Article Snippet: Then, the membranes were incubated overnight at 4 °C with 1:1000 dilutions of antibodies against TRIM27 (Proteintech, #12205-1-AP), VCAM-1 (Abcam, #ab134047, RRID: AB_2721053), Janus kinase 2 (JAK2; Proteintech, #17670-1-AP, RRID: AB_2811138), p-JAK2 (Y1007) (Abcam, #ab195055), signal transducer and activator of transcription 3 (STAT3; Proteintech, #60199-1-Ig, RRID: AB_10913811), p-STAT3 (Tyr705) (CST, #9145, RRID: AB_2491009), nephrin (Abcam, #ab58968, RRID: AB_944400), podocin (Abcam, #ab50339, RRID: AB_882097), CD63 (Santa Cruz Biotechnology, Dallas, TX, USA, #sc-5275, RRID: AB_627877), CD9 (Santa Cruz Biotechnology, #sc-13118, RRID: AB_627213), TSG101 (Proteintech, #28283-1-AP, RRID: AB_2881104), Calnexin (Proteintech, #10427-2-AP, RRID: AB_2069033), Rab27a (Proteintech, #17817-1-AP, RRID: AB_2176728), PTEN (Abcam, #ab32199, RRID: AB_777535), Akt (Proteintech, #60203-2-Ig, RRID: AB_10912803), and p-Akt (S473) (Abcam, #ab81283, RRID: AB_2224551).

Techniques: Western Blot, Expressing, Control, Enzyme-linked Immunosorbent Assay, Knockdown

Journal: Cell Death Discovery

Article Title: TRIM27-controlled endothelium-derived exosomes play a central role in podocyte injury in diabetic kidney disease

doi: 10.1038/s41420-026-02953-y

Figure Lengend Snippet: A – D Western blot assay showed p-JAK2 (Y1007) and p-STAT3 (Tyr705) expression increased in HRGECs treated with HG or TGF-β1. ** P < 0.01, *** P < 0.001, **** P < 0.0001 vs. 0 h group ( n = 3). E , F Western blot assay showed p-JAK2 (Y1007), p-STAT3 (Tyr705), and VCAM-1 expression decreased after inhibition of the JAK2/STAT3 pathway by AG490 in HRGECs and treatment with HG for 24 h. ** P < 0.01, *** P < 0.001, **** P < 0.0001 vs. control group, ## P < 0.01 vs. HG + DMSO group ( n = 3). G – I IF assay showed that VE-cadherin, ZO-1, and syndecan-1 expression increased in HRGECs treated with AG490 and TGF-β1 for 24 h. Scale bars: 25 μm. J , K The ELISA assays showed contents of syndecan-1 and VCAM-1 in culture supernatants of HRGECs decreased after treated with AG490 and HG for 24 h. * P < 0.05, **** P < 0.0001 vs. control group, # P < 0.05, ### P < 0.001 vs. HG + DMSO group ( n = 3). L , M Western blot showed that VCAM-1 expression decreased after inhibition of the JAK2/STAT3 pathway in HRGECs. ** P < 0.01 vs. control group, ### P < 0.001 vs. TGF-β1 + DMSO group ( n = 3). N , O Western blot assay showed downregulation of TRIM27 decreased p-JAK2 (Y1007) and p-STAT3 (Tyr705) expression in HRGECs treated with HG for 2 h. *** P < 0.001, **** P < 0.0001 vs. control group, ### P < 0.001 vs. HG+shNC group ( n = 3). Bonferroni’s correction was performed to analyze statistical significance. Data are the mean ± SEM.

Article Snippet: Then, the membranes were incubated overnight at 4 °C with 1:1000 dilutions of antibodies against TRIM27 (Proteintech, #12205-1-AP), VCAM-1 (Abcam, #ab134047, RRID: AB_2721053), Janus kinase 2 (JAK2; Proteintech, #17670-1-AP, RRID: AB_2811138), p-JAK2 (Y1007) (Abcam, #ab195055), signal transducer and activator of transcription 3 (STAT3; Proteintech, #60199-1-Ig, RRID: AB_10913811), p-STAT3 (Tyr705) (CST, #9145, RRID: AB_2491009), nephrin (Abcam, #ab58968, RRID: AB_944400), podocin (Abcam, #ab50339, RRID: AB_882097), CD63 (Santa Cruz Biotechnology, Dallas, TX, USA, #sc-5275, RRID: AB_627877), CD9 (Santa Cruz Biotechnology, #sc-13118, RRID: AB_627213), TSG101 (Proteintech, #28283-1-AP, RRID: AB_2881104), Calnexin (Proteintech, #10427-2-AP, RRID: AB_2069033), Rab27a (Proteintech, #17817-1-AP, RRID: AB_2176728), PTEN (Abcam, #ab32199, RRID: AB_777535), Akt (Proteintech, #60203-2-Ig, RRID: AB_10912803), and p-Akt (S473) (Abcam, #ab81283, RRID: AB_2224551).

Techniques: Western Blot, Expressing, Inhibition, Control, Enzyme-linked Immunosorbent Assay

Journal: Cell Death Discovery

Article Title: TRIM27-controlled endothelium-derived exosomes play a central role in podocyte injury in diabetic kidney disease

doi: 10.1038/s41420-026-02953-y

Figure Lengend Snippet: A Eighteen 20-week-old mice were randomly divided into three groups: STZ ( n = 6), STZ+shTRIM27 ( n = 6), and STZ+shNC group ( n = 6). Mice in STZ+shTRIM27 and STZ+shNC groups were renally injected with 50 μl of 1 × 10 11 infective units of adeno-associated virus at three sites each in both kidneys. Six control mice and 6 STZ mice were injected with isometric saline. The mice were sacrificed after 4 weeks. B IF staining showed TRIM27 expression decreased in glomerular endothelial cells of STZ+shTRIM27 mice. Scale bars: 10 μm. C – E Level of 24-h proteinuria, BUN, and Scr decreased in STZ+shTRIM27 mice. ** P < 0.01, *** P < 0.001 vs. Control mice, ## P < 0.01 vs. STZ+shNC mice. n = 6 each group. F , G The ELISA assays showed serum contents of syndecan-1 and VCAM-1 decreased in STZ+shTRIM27 mice. * P < 0.05, *** P < 0.001 vs. Control mice, # P < 0.05, ## P < 0.01 vs. STZ+shNC mice. n = 6 each group. H , I IF staining showed syndecan-1 expression increased in STZ+shTRIM27 mice. Scale bars: 25 μm. *** P < 0.001 vs. Control mice, ### P < 0.001 vs. STZ+shNC mice ( n = 6). J Electron microscopy assay showed ultrastructure of the podocytes in the renal cortex of mice. Scale bars: 2 μm. K – M IHC staining showed WT1 and podocin expression increased in STZ+shTRIM27 mice. Scale bars: 25 μm. *** P < 0.001 vs. Control mice, ## P < 0.01 vs. STZ+shNC mice ( n = 6). N IF staining showed synaptopodin expression increased in STZ+shTRIM27 mice. Scale bars: 10 μm. Bonferroni’s correction was performed to analyze statistical significance. Data are the mean ± SEM.

Article Snippet: Then, the membranes were incubated overnight at 4 °C with 1:1000 dilutions of antibodies against TRIM27 (Proteintech, #12205-1-AP), VCAM-1 (Abcam, #ab134047, RRID: AB_2721053), Janus kinase 2 (JAK2; Proteintech, #17670-1-AP, RRID: AB_2811138), p-JAK2 (Y1007) (Abcam, #ab195055), signal transducer and activator of transcription 3 (STAT3; Proteintech, #60199-1-Ig, RRID: AB_10913811), p-STAT3 (Tyr705) (CST, #9145, RRID: AB_2491009), nephrin (Abcam, #ab58968, RRID: AB_944400), podocin (Abcam, #ab50339, RRID: AB_882097), CD63 (Santa Cruz Biotechnology, Dallas, TX, USA, #sc-5275, RRID: AB_627877), CD9 (Santa Cruz Biotechnology, #sc-13118, RRID: AB_627213), TSG101 (Proteintech, #28283-1-AP, RRID: AB_2881104), Calnexin (Proteintech, #10427-2-AP, RRID: AB_2069033), Rab27a (Proteintech, #17817-1-AP, RRID: AB_2176728), PTEN (Abcam, #ab32199, RRID: AB_777535), Akt (Proteintech, #60203-2-Ig, RRID: AB_10912803), and p-Akt (S473) (Abcam, #ab81283, RRID: AB_2224551).

Techniques: Injection, Virus, Control, Saline, Staining, Expressing, Enzyme-linked Immunosorbent Assay, Electron Microscopy, Immunohistochemistry

Journal: Cell Death Discovery

Article Title: TRIM27-controlled endothelium-derived exosomes play a central role in podocyte injury in diabetic kidney disease

doi: 10.1038/s41420-026-02953-y

Figure Lengend Snippet: A , B Concentrations and size distributions of T+shTRIM27-exo and T+shNC-exo determined by NTA. C Quantification of NTA results from three independent experiments. ** P < 0.01 vs. T+shTRIM27 group ( n = 3). D – G Western blot assay showed downregulation of TRIM27 decreased Rab27a expression in HRGECs treated with TGF-β1 or HG for 24 h. * P < 0.05, *** P < 0.001 vs. control group, # P < 0.05, ## P < 0.01 vs. TGF-β1+shNC or HG+shNC group ( n = 3). H – K Western blot assay showed conditional media from HRGECs with knocked down TRIM27 increased nephrin and podocin expression in HPCs. * P < 0.05, *** P < 0.001 vs. CM group, ## P < 0.01, ### P < 0.001 vs. HM+shNC group or TM+shNC group ( n = 3). L Table of the 10 most abundant miRNAs in HG exosomes compared with control exosomes. M – P Detection of miR-486-5p in HRGEC-derived exosomes and HPCs treated with exosomes by qPCR normalized to U6. **** P < 0.0001 vs. C-exo group, ns. no significance ( n = 6). Student’s t test and Bonferroni’s correction were performed to analyze statistical significance. Values are the mean ± SEM.

Article Snippet: Then, the membranes were incubated overnight at 4 °C with 1:1000 dilutions of antibodies against TRIM27 (Proteintech, #12205-1-AP), VCAM-1 (Abcam, #ab134047, RRID: AB_2721053), Janus kinase 2 (JAK2; Proteintech, #17670-1-AP, RRID: AB_2811138), p-JAK2 (Y1007) (Abcam, #ab195055), signal transducer and activator of transcription 3 (STAT3; Proteintech, #60199-1-Ig, RRID: AB_10913811), p-STAT3 (Tyr705) (CST, #9145, RRID: AB_2491009), nephrin (Abcam, #ab58968, RRID: AB_944400), podocin (Abcam, #ab50339, RRID: AB_882097), CD63 (Santa Cruz Biotechnology, Dallas, TX, USA, #sc-5275, RRID: AB_627877), CD9 (Santa Cruz Biotechnology, #sc-13118, RRID: AB_627213), TSG101 (Proteintech, #28283-1-AP, RRID: AB_2881104), Calnexin (Proteintech, #10427-2-AP, RRID: AB_2069033), Rab27a (Proteintech, #17817-1-AP, RRID: AB_2176728), PTEN (Abcam, #ab32199, RRID: AB_777535), Akt (Proteintech, #60203-2-Ig, RRID: AB_10912803), and p-Akt (S473) (Abcam, #ab81283, RRID: AB_2224551).

Techniques: Western Blot, Expressing, Control, Derivative Assay