rab7  (OriGene)

Journal: Journal of Nanobiotechnology

Article Title: Activation of the Wnt/β-catenin signalling pathway enhances exosome production by hucMSCs and improves their capability to promote diabetic wound healing

doi: 10.1186/s12951-024-02650-x

Figure Lengend Snippet: CHIR99021 increased exosome secretion by promoting SNAP25-dependent exocytosis. ( A ) Volcano plot showing differential gene expression of hucMSCs in the CHIR99021-treated group versus the control group. ( B ) GO enrichment analysis of hucMSCs in CHIR99021-treated group versus control group. ( C ) KEGG enrichment analysis of hucMSCs in CHIR99021-treated group versus control group. ( D ) Heatmap of gene clustering of hucMSCs transcriptomics in synaptic vesicle cycle between CHIR99021-treated and control groups. ( E ) Western blot analysis of β-catenin, Rab27a, Rab7, SNAP25 in hucMSCs treated with Wnt signaling pathway agonists and inhibitors. ( F ) Co-location of SNAP25 with VAMP3 was observed using immunofluorescence after treatment of hucMSCs with Wnt signaling pathway agonists and inhibitors. ( G ) Co-location of LAMP2 with CD63 was observed using immunofluorescence after treatment of hucMSCs with Wnt signaling pathway agonists and inhibitors. ( H ) Western blot analysis of SNAP25 knockdown efficiency. ( I ) After knocking down of SNAP25, hucMSCs were treated with Wnt signaling pathway agonists for 48 h, supernatants were collected, and AChE concentration was analysis by ELISA. Data are represented as mean ± SEM. * P < 0.05, ** P < 0.01

Article Snippet: Primary antibodies against the following target proteins were used: SNAP25 (1:100; Proteintech 60159-1-Ig), VAMP3 (1:100; Proteintech 10702-1-AP), LAMP2 (1:200; ABclonal a22482), CD63 (1:200; Proteintech 25682-1-AP), and Rab7a (1:200; ABclonal a12308).

Techniques: Gene Expression, Control, Western Blot, Immunofluorescence, Knockdown, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Animal Nutrition

Article Title: Retinoic acid alleviates rotavirus-induced intestinal damage by regulating redox homeostasis and autophagic flux in piglets

doi: 10.1016/j.aninu.2023.12.007

Figure Lengend Snippet: Effects of retinoic acid (RA) administration on autophagy activation in jejunum from control and RA-treated groups. (A and B) Western blotting analysis of autophagy-related proteins including autophagy induction and regulation (pBeclin/Beclin, pULK1/ULK1, ATG7, ATG16 and ATG14), autophagy cargo recognition and degradation (p62, ATG5, ATG3), and autophagy maturation and lysosomal fusion (syntaxin, SNAP29, Rab7A and LC3I/II) ( n = 3). (C to G) Gray value analysis of the above protein expressions ( n = 3). KD = kiloDalton; Beclin = coiled-coil, moesin-like BCL2-interacting protein; ULK1 = Unc51-like kinase 1; ATG = autophagy-related gene; p62 = sequestosome 1; SNAP29 = synaptosome associated protein 29; RAB7A = RAS-related protein Rab-7a; LC3 = microtubule-associated protein light chain 3. ∗ P < 0.05, ∗∗ P < 0.01 and ∗∗∗ P < 0.001.

Article Snippet: Western blotting was conducted as previously described ( ) using primary antibodies against zonula occludens-1 (ZO-1) (13663, Cell Signaling Technology, Danvers, MA, USA), occludin (911131, Cell Signaling Technology, Danvers, MA, USA), claudin-1 (13255, Cell Signaling Technology, Danvers, MA, USA), pNRF2 (ab76026, Abcam, Cambridge, UK), NRF2 (16396-1-AP, Proteintech, Wuhan, China), heme oxygenase 1 (HO-1) (10701-1-AP, Proteintech, Wuhan, China), sequestosome 1 (p62) (ab56416, Abcam, Cambridge, UK), glutathione peroxidase 4 (GPX4) (ab125066, Abcam, Cambridge, UK), Unc51-like kinase 1 (ULK1) (8054, Cell Signaling Technology, Danvers, MA, USA), pULK1 (14202, Cell Signaling Technology, Danvers, MA, USA), autophagy-related gene (ATG) 7 (8558, Cell Signaling Technology, Danvers, MA, USA), ATG16 (8089, Cell Signaling Technology, Danvers, MA, USA), ATG14 (96752, Cell Signaling Technology, Danvers, MA, USA), coiled-coil, moesin-like BCL2-interacting protein (Beclin) (3495, Cell Signaling Technology, Danvers, MA, USA), pBeclin (14717, Cell Signaling Technology, Danvers, MA, USA), ATG5 (9980, Cell Signaling Technology, Danvers, MA, USA), ATG3 (3415, Cell Signaling Technology, Danvers, MA, USA), syntaxin (2869, Cell Signaling Technology, Danvers, MA, USA), synaptosome associated protein 29 (SNAP29) (sc-390602, Santa Cruz, Dallas, Texas, USA), RAS-related protein Rab-7a (RAB7A) (9367, Cell Signaling Technology, Danvers, MA, USA), LC3I/II (4108, Cell Signaling Technology, Danvers, MA, USA), and β-actin (3700, Cell Signaling Technology, Danvers, MA, USA).

Techniques: Activation Assay, Western Blot

Journal: Animal Nutrition

Article Title: Retinoic acid alleviates rotavirus-induced intestinal damage by regulating redox homeostasis and autophagic flux in piglets

doi: 10.1016/j.aninu.2023.12.007

Figure Lengend Snippet: Effects of retinoic acid (RA) administration on regulating autophagic pathways in jejunum from control, rotavirus (RV) and RV + RA groups. (A) Fluorescence density of RV in all six treatment groups in jejunum sample (red fluorescence indicates RV protein detection). Scale bar = 200 μm. (B and C) Western blotting analysis of autophagy-related proteins including autophagy induction and regulation (pBeclin/Beclin, pULK1/ULK1, ATG7, ATG16 and ATG14), autophagy cargo recognition and degradation (p62, ATG5 and ATG3), and autophagy maturation and lysosomal fusion (syntaxin, SNAP29, RAB7A and LC3I/II) ( n = 3). (D to I) Gray value analysis of the above protein expressions ( n = 3). KD = kiloDalton; Beclin = coiled-coil, moesin-like BCL2-interacting protein; ULK1 = Unc51-like kinase 1; ATG = autophagy-related gene; p62 = sequestosome 1; SNAP29 = synaptosome associated protein 29; RAB7A = RAS-related protein Rab-7a; LC3 = microtubule-associated protein light chain 3. ∗ P < 0.05, and ∗∗ P < 0.01.

Article Snippet: Western blotting was conducted as previously described ( ) using primary antibodies against zonula occludens-1 (ZO-1) (13663, Cell Signaling Technology, Danvers, MA, USA), occludin (911131, Cell Signaling Technology, Danvers, MA, USA), claudin-1 (13255, Cell Signaling Technology, Danvers, MA, USA), pNRF2 (ab76026, Abcam, Cambridge, UK), NRF2 (16396-1-AP, Proteintech, Wuhan, China), heme oxygenase 1 (HO-1) (10701-1-AP, Proteintech, Wuhan, China), sequestosome 1 (p62) (ab56416, Abcam, Cambridge, UK), glutathione peroxidase 4 (GPX4) (ab125066, Abcam, Cambridge, UK), Unc51-like kinase 1 (ULK1) (8054, Cell Signaling Technology, Danvers, MA, USA), pULK1 (14202, Cell Signaling Technology, Danvers, MA, USA), autophagy-related gene (ATG) 7 (8558, Cell Signaling Technology, Danvers, MA, USA), ATG16 (8089, Cell Signaling Technology, Danvers, MA, USA), ATG14 (96752, Cell Signaling Technology, Danvers, MA, USA), coiled-coil, moesin-like BCL2-interacting protein (Beclin) (3495, Cell Signaling Technology, Danvers, MA, USA), pBeclin (14717, Cell Signaling Technology, Danvers, MA, USA), ATG5 (9980, Cell Signaling Technology, Danvers, MA, USA), ATG3 (3415, Cell Signaling Technology, Danvers, MA, USA), syntaxin (2869, Cell Signaling Technology, Danvers, MA, USA), synaptosome associated protein 29 (SNAP29) (sc-390602, Santa Cruz, Dallas, Texas, USA), RAS-related protein Rab-7a (RAB7A) (9367, Cell Signaling Technology, Danvers, MA, USA), LC3I/II (4108, Cell Signaling Technology, Danvers, MA, USA), and β-actin (3700, Cell Signaling Technology, Danvers, MA, USA).

Techniques: Fluorescence, Western Blot