protein kinase array  (R&D Systems)

Journal: Stem cell research & therapy

Article Title: Restoration of NOX4 signalling reverses endothelial colony-forming cell angiogenic dysfunction associated with experimental and clinical diabetes.

doi: 10.1186/s13287-025-04393-4

Figure Lengend Snippet: Fig. 6 Clinical diabetes induces dysregulation of CB-ECFC angiogenic pathways linked with downstream NOX4-dependent pro-angiogenic signalling. CB-ECFCs were isolated from healthy and gestational diabetic donors and subjected to RNA sequencing prior to (A) IPA network generation, node colour represents predicted gene activation (orange) and inhibition (blue). B, C Proteome Profiler™ analysis of pooled lysates (triplicates from 5 clones) from diabetic CB-ECFCs subjected to electroporation for introduction of either empty vector (EV; pcDNA4/TO/myc-His A) or NOX4 overexpression (OE; pcDNA4/TO/NOX4-myc-His A) construct for detection of protein phosphorylation. Differential expression between groups presented as a heat map indicating largely decreased (red) phosphorylation (OE versus EV). D–G Protein expression of endoglin, E2F1 and SERPINE1 by Western blotting with normalisation to ACTB as loading control; n = 8/9, combined data from 3 different clones. Representative cropped blots shown from one healthy and one diabetic clone. Full-length blots/gels are presented in Supplementary Figure. Data are mean ± SEM; **P < 0.01, ***P < 0.001, paired Student’s t-test

Article Snippet: Proteome profiler arrays (R&D Systems) specific for protein phosphorylation (Human phospho-kinase proteome profiler array, ARY003C) or linked with angiogenesis (Human angiogenesis proteome profiler array; ARY007) were run according to manufacturer’s instructions.

Techniques: Isolation, RNA Sequencing, Activation Assay, Inhibition, Clone Assay, Electroporation, Plasmid Preparation, Over Expression, Construct, Phospho-proteomics, Quantitative Proteomics, Expressing, Western Blot, Control

Journal: Stem cell research & therapy

Article Title: Restoration of NOX4 signalling reverses endothelial colony-forming cell angiogenic dysfunction associated with experimental and clinical diabetes.

doi: 10.1186/s13287-025-04393-4

Figure Lengend Snippet: Fig. 7 Restoration of NOX4 expression in diabetic CB-ECFCs rescues angiogenic capacity by specifically inducing pro-angiogenic signalling. Summary schematic indicating that restoration of NOX4 levels in diabetic CB-ECFCs leads to upregulated E2F1 signalling in parallel with fully restored angiogenic function. E2F1 is a transcription factor which positively regulates key genes associated with efficient progression through the cell cycle, DNA replication and subsequent proliferation, which is critical to support an efficient and potent pro-angiogenic response. NOX4 induction in diabetic CB-ECFCs also led to reduced phosphorylation of anti-proliferative P53 (at S46), increased expression of SERPINE1 and endoglin, which are implicated in pro-angiogenic signalling

Article Snippet: Proteome profiler arrays (R&D Systems) specific for protein phosphorylation (Human phospho-kinase proteome profiler array, ARY003C) or linked with angiogenesis (Human angiogenesis proteome profiler array; ARY007) were run according to manufacturer’s instructions.

Techniques: Expressing, Phospho-proteomics