gapdh proteintech group  (Proteintech)

Journal: Cell death & disease

Article Title: RNA N 6 -methyladenosine demethylase FTO promotes diabetic wound healing through TRIB3-mediated autophagy in an m 6 A-YTHDF2-dependent manner.

doi: 10.1038/s41419-025-07494-3

Figure Lengend Snippet: Fig. 6 FTO influenced proliferation and migration by affecting the expression of TRIB3. A-D Overexpression of TRIB3 rescued the proliferative activity of HaCaT and NHEK cells with FTO knockout, as reflected by CCK-8 and colony formation assays. Semi-quantitative analysis of colony formation assays is shown (n = 3). E, F The Edu incorporation assays revealed that FTO significantly inhibits proliferation of keratinocytes, which can be rescued by overexpression of TRIB3. G, H Overexpression of TRIB3 during FTO knockdown partially rescued the migration of keratinocytes. Images were captured at 0 h and 12 h after the scratch. Semi-quantitative analysis of wound-healing assays is shown (n = 3). I Timeline of in vivo experiments for STZ injections, sgRNA and vector injections, and observation of wound healing models in mice. J Measurements of blood glucose levels in STZ-treated mice. K-M Representative images of cutaneous wounds of diabetic mice, diabetic mice with koFTO injections, diabetic mice with oeTRIB3 injections, and diabetic mice with koFTO and oeTRIB3 injections on days 0, 4, 8, and 12 after wound generation by surgical excision. Rates of wound closure were quantified using ImageJ software and expressed as the percentage of closed wound area (n = 5 per group). The data are presented as the mean ± SD or mean ± SEM (A, B, D, F, H, J, M), and P-values of all data by a two-tailed unpaired t-test are indicated. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: Target Application Supplier Catalog Number FTO Western blot/IHC/IF Abcam ab126605 FTO Western blot/IHC/IF Novus NBP1-77021 m6A Dot blot/IF Synaptic Systems 202 003 cytokeratin IF ORIGENE BP5069 SQSTM1 Western blot/CO-IP/IF CST #23214 LC3A/B Western blot/IHC CST #12741 TRIB3 CO-IP Proteintech Group 13300-1-AP TRIB3 Western blot CST #43043 YTHDF2 Western blot/ CO-IP Proteintech Group 24744-1-AP HA-Tag Western blot Abmart M20003 ATG5 Western blot Abmart T55766 ATG7 Western blot Abmart T55658 ULK1 Western blot Abmart T56902 IgG RIP/Co-IP Abcam ab172730 GAPDH Western blot Proteintech Group 60004-1-Ig ACTB Western blot Proteintech Group 60008-1-Ig Table.

Techniques: Migration, Expressing, Over Expression, Activity Assay, Knock-Out, CCK-8 Assay, Knockdown, In Vivo, Plasmid Preparation, Software, Two Tailed Test

Journal: Cell death & disease

Article Title: RNA N 6 -methyladenosine demethylase FTO promotes diabetic wound healing through TRIB3-mediated autophagy in an m 6 A-YTHDF2-dependent manner.

doi: 10.1038/s41419-025-07494-3

Figure Lengend Snippet: Fig. 7 FTO regulated autophagy by targeting TRIB3, which inhibited autophagic degradation by interacting with SQSTM1. A, B TRIB3, SQSTM1, LC3I, and LC3II protein levels were measured by western blot in HaCaT and NHEK cells transfected with lentiviruses carrying koFTO and/or oeTRIB3 (n = 3). C–E The HaCaT and NHEK cells were infected with adenovirus harboring tandem fluorescent mRFP-GFP-LC3 for 24 h, followed by transfection with oeTRIB3, koFTO, and koFTO with oeTRIB3 in normal-glucose culture medium. Representative images of the HaCaT and NHEK cells expressing mRFP-GFP-LC3 are shown. Green: GFP puncta; red: mRFP puncta. Semi-quantitative analysis of autophagosomes (AP, yellow puncta in merged images) and autolysosomes (AL, red-only puncta in merged images, n = 10 randomly selected conditions from 3 independent experiments, Scale bar, 50 μm). F The TRIB3-SQSTM1 interaction was evaluated by an IP assay in TRIB3- overexpressing HaCaT cells (n = 3). G, I Colocalization of TRIB3 and SQSTM1 was detected by immunostaining in control and TRIB3- overexpressing HaCaT and NHEK cells. H, J Line chart of fluorescence signal positioning analysis (n = 3; Scale bar, 10 μm). K–M Expression of TRIB3 and colocalization of TRIB3 with SQSTM1 in Con and DW tissues were detected by immunostaining. (n = 5 per group; Scale bar, 50 μm). The data are presented as the mean ± SD or mean ± SEM (B, D, E), and P-values of all data by a two-tailed unpaired t-test are indicated. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: Target Application Supplier Catalog Number FTO Western blot/IHC/IF Abcam ab126605 FTO Western blot/IHC/IF Novus NBP1-77021 m6A Dot blot/IF Synaptic Systems 202 003 cytokeratin IF ORIGENE BP5069 SQSTM1 Western blot/CO-IP/IF CST #23214 LC3A/B Western blot/IHC CST #12741 TRIB3 CO-IP Proteintech Group 13300-1-AP TRIB3 Western blot CST #43043 YTHDF2 Western blot/ CO-IP Proteintech Group 24744-1-AP HA-Tag Western blot Abmart M20003 ATG5 Western blot Abmart T55766 ATG7 Western blot Abmart T55658 ULK1 Western blot Abmart T56902 IgG RIP/Co-IP Abcam ab172730 GAPDH Western blot Proteintech Group 60004-1-Ig ACTB Western blot Proteintech Group 60008-1-Ig Table.

Techniques: Western Blot, Transfection, Infection, Expressing, Immunostaining, Control, Two Tailed Test

Journal: Cell death & disease

Article Title: RNA N 6 -methyladenosine demethylase FTO promotes diabetic wound healing through TRIB3-mediated autophagy in an m 6 A-YTHDF2-dependent manner.

doi: 10.1038/s41419-025-07494-3

Figure Lengend Snippet: Fig. 8 Proposed working models of FTO in the regulation of autophagy in keratinocytes of diabetic skin. The expression of TRIB3 was regulated by FTO, and it interacted and cooperated with SQSTM1 to induce autophagy in keratinocytes. In diabetes, downregulation of FTO induced decreased expression levels of TRIB3 via acceleration of TRIB3 mRNA decay, which led to the inhibition of autophagy and the impairment of keratinocyte migration, eventually resulting in delayed wound healing.

Article Snippet: Target Application Supplier Catalog Number FTO Western blot/IHC/IF Abcam ab126605 FTO Western blot/IHC/IF Novus NBP1-77021 m6A Dot blot/IF Synaptic Systems 202 003 cytokeratin IF ORIGENE BP5069 SQSTM1 Western blot/CO-IP/IF CST #23214 LC3A/B Western blot/IHC CST #12741 TRIB3 CO-IP Proteintech Group 13300-1-AP TRIB3 Western blot CST #43043 YTHDF2 Western blot/ CO-IP Proteintech Group 24744-1-AP HA-Tag Western blot Abmart M20003 ATG5 Western blot Abmart T55766 ATG7 Western blot Abmart T55658 ULK1 Western blot Abmart T56902 IgG RIP/Co-IP Abcam ab172730 GAPDH Western blot Proteintech Group 60004-1-Ig ACTB Western blot Proteintech Group 60008-1-Ig Table.

Techniques: Expressing, Inhibition, Migration

Journal: iScience

Article Title: SIRT3 differentially regulates lysine benzoylation from SIRT2 in mammalian cells

doi: 10.1016/j.isci.2024.111176

Figure Lengend Snippet:

Article Snippet: GAPDH , Proteintech Group , 60004-1-Ig; RRID: AB_2107436.

Techniques: Virus, Recombinant, Protease Inhibitor, Transfection, Kinase Assay, shRNA, Sequencing, Plasmid Preparation, Modification, Software