Journal: Cell stem cell

Article Title: hPSC-derived sacral neural crest enables rescue in a severe model of Hirschsprung's disease.

doi: 10.1016/j.stem.2023.02.003

Figure Lengend Snippet: Figure 5. Vagal and sacral NC exhibits distinct behavior in vitro (A) Schematic drawing of experimental design. RFP-tagged hPSC line was used for vagal lineages and GFP-tagged line for sacral lineages. (B) qRT-PCR of SOX10 and HOX genes to confirm the VNC and SNC identity. n = 6 biological replicates.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Goat anti SOX10 Santa Cruz Cat#sc-17342; RRID:AB_2195374 Mouse anti SOX10 Santa Cruz Cat#sc-365692; RRID:AB_10844002 Mouse anti HOXC9 Abcam Cat#ab50839; RRID:AB_880494 Rabbit anti HOXD13 Abcam Cat#ab19866; RRID:AB_733004 Mouse anti Ki67 Dako Cat# M7240; RRID:AB_2142367 Mouse anti CDX2 BioGenex Cat#MU392A-5UC; RRID:AB_2650531 Goat anti Bra/T R&D Systems Cat#AF2085; RRID:AB_2200235 Rabbit anti SOX2 Cell signaling Tech Cat#35795; RRID:AB_2195767 PE conjugated Mouse Anti-Human CDX-2 BD Biosciences Cat#563428; RRID:AB_2738198) APC-conjugated Human/Mouse Bra/T R&D Systems Cat#IC2085A; RRID:AB_2891298 Alexa Fluor 488 Mouse anti-Sox2 Clone O30-678 BD Biosciences Cat#561593; RRID:AB_10894382 Rabbit anti SNAI2 Cell signaling Tech Cat#9585; RRID:AB_2239535 Rabbit anti TUJ1 Covance Cat#MMS-435P; RRID:AB_2313773 Chicken anti TUJ1 Abcam Cat#ab107216; RRID:AB_10899689 Mouse anti TUJ1 Biolegend Cat#801201; RRID:AB_2313773 Rabbit anti GABA Sigma Cat#A2052; RRID:AB_477652 Mouse anti TH Immunostar Cat#22941; RRID:AB_572268 Rabbit anti nNOS Millipore Cat#07-571; RRID:AB_11211970 Goat anti CHAT Millipore Cat#AB144P; RRID:AB_2079751 Rabbit anti 5’HT Sigma Cat#S5545; RRID:AB_477522 Rabbit anti SST Millipore Cat#mab354; RRID:AB_2255365 Rabbit anti DBH ImmunoStar Cat#22806; RRID:AB_572229 Rabbit ISL1 Developmental Studies Hybridoma Bank Cat#39.4D5; RRID:AB_2314683 Rabbit anti S100-beta Abcam Cat# ab52642; RRID:AB_882426 PE/Cy7 conjugated CD49D Biolegend Cat#304314; RRID:AB_10643278 APC conjugated CD49D Biolegend Cat#304308; RRID:AB_2130041 PE conjugated HNK1 Biolegend Cat#359611; RRID:AB_2562758 APC conjugated P75NTR Biolegend Cat#345107; RRID:AB_10639737 APC conjugated c-kit (CD117) eBioscience Cat#17-1179-42; RRID:AB_10596820 Chicken anti GFP Abcam Cat#ab13970; RRID:AB_300798 Rabbit anti RFP Rockland Cat#600-401-379; RRID:AB_2209751 Mouse anti Stem121 TaKaRa Cat#Y40410; RRID:AB_2801314 Mouse anti RET R&D systems Cat#MAB718-100; RRID:AB_2232594 Rabbit anti EDNRB Biorbyt Cat#orb37019; RRID:AB_10995764 Anti-rabbit IgG, HRP-linked Cell Signaling Tech Cat#7074S; RRID:AB_2099233 Rabbit anti LaminB1 Abcam Cat#ab16048; RRID:AB_443298 Human anti ANNA-1 Gift from Dr V. Lennon (King et al., 1999) Donkey anti-Human IgG H&L (AMCA) Jackson Immuno Cat# 709-155-149; RRID: AB_2340529 Donkey anti-Human Dylight 405 Jackson Immuno Cat# 709-475-149; RRID: AB_2340553 Donkey anti-goat IgG H&L (Alexa Fluor 488) Invitrogen Cat#A11055; RRID:AB_2534102 (Continued on next page) Cell Stem Cell 30, 264–282.e1–e9, March 2, 2023 e1

Techniques: In Vitro, Quantitative RT-PCR

Journal: Heliyon

Article Title: Median raphe glutamatergic neuron-mediated enhancement of GABAergic transmission and suppression of long-term potentiation in the hippocampus

doi: 10.1016/j.heliyon.2024.e38192

Figure Lengend Snippet: MRN VGLUT3 neuron-mediated glutamatergic transmission in SR/SLM 5-HT3aR neurons. A) A schematic of AAV injection into the MRN. The left panel shows an image of the MRN from VGLUT3 Cre/5-HT3aR-EGFP mouse which received an AAV-EF1a-double floxed-hChR2(H134R)-mCherry-WPRE-HGHpA injection into the MRN. Scale 150 μm. B) An image of the hippocampus from MRN VGLUT3 ChR2/5-HT3aR-EGFP mouse. The inset shows EGFP-expressing SR/SLM 5-HT3aR neurons and mCherry-expressing MRN VGLUT3 axons. Scale 300 μm/25 μm (inset). C) The upper panel shows a schematic for recording MRN VGLUT3 neuron-mediated transmission in SR/SLM 5-HT3aR neurons. The bottom panel shows example traces for MRN VGLUT3 neuron-mediated EPSCs in SR/SLM 5-HT3aR neurons without drug application, in the presence of tetrodotoxin (TTX, 1 μM), TTX+4-aminopyridine (4-AP, 100 μM) and TTX+4-AP + DNQX (10 μM). Scale 50 pA/10 ms. The blue line indicates light application. D) EPSC amplitude in SR/SLM 5-HT3aR neurons before the drug application, in the presence of TTX, TTX+4-AP, and TTX+4-AP + DNQX (6 neurons from 3 female mice and 6 neurons from 3 male mice). F (1,11) = 22.01, P = 0.001. Data are presented as mean ± SEM. E) EPSC amplitude in female (17 neurons/5 mice) and male (16 neurons/5 mice) groups. Female and male groups did not show statistically significant difference in EPSC amplitude (P = 0.8). The horizontal line in each group represents the mean and the vertical line represents SEM. F) The delay between the onset of the light stimulation and the onset of EPSCs in female (17 neurons/5 mice) and male (16 neurons/5 mice) groups. Female and male groups did not show a statistically significant difference in the delay between the onset of light stimulation and the onset of EPSCs (P = 0.77). n.s. not statistically significant.

Article Snippet: The slices were then incubated in mCherry polyclonal antibody (Rockland Immunochemicals, Cat. No. 600401P16) or Anti-GFP antibody (Abcam, Cat. No. ab13970, RRID: AB_300798 ) at a dilution of 1:1000 in 0.3 % Triton-X/TBS for 24 h at 4 °C [ , ].

Techniques: Transmission Assay, Injection, Expressing

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