Journal: The Journal of Biological Chemistry

Article Title: A tandem recruitment site in the pseudokinase scaffold PEAK3 is subject to phosphorylation-dependent regulation and cancer-associated mutations

doi: 10.1016/j.jbc.2026.111365

Figure Lengend Snippet: Serine/threonine protein kinases regulating PEAK3 S69 phosphorylation. A , consensus motifs of serine/threonine protein kinases with selectivity corresponding to the PEAK3 S69 region. Phosphorylation sites (position 0) are highlighted in pink and key features highlighted are arginine or lysine at position −3 ( orange ) and hydrophobic residues (ϕ) at +1 and −5 position ( blue ). X = any amino acid. Motifs sourced from Johnson et al. and PhosphoSitePlus ( , ). B , role of PKCs. MCF-10A/PEAK3 cells were treated with Go6850 or Go6976 and cell lysates subjected to Western blotting as indicated. Results are representative of four independent experiments. Histogram indicates the relative level of PEAK3 p-S69 normalized for HA and expressed relative to the DMSO control, which was arbitrarily set to 1.0. Data represent mean ± S.E.M., with 'au' indicating arbitrary units. ∗ p < 0.05, ∗∗ p < 0.01, by ratio paired t test. C , role of CaMKII as determined by pharmacological inhibition. MCF-10A/PEAK3 cells were treated with RA306 or vehicle control and cell lysates analyzed by Western blotting as indicated. Data are representative of three independent experiments. Results in C are derived from the same experiment and use the same DMSO control. Histogram indicates the relative level of PEAK3 p-S69 as mean ± S.E.M., ∗∗ p < 0.01, by ratio paired t test. D , role of CaMKII as determined by knockdown. CaMKIID and G were subject to siRNA-mediated knockdown (KD) in MCF-10A/PEAK3 cells and PEAK3 p-S69 assayed by Western blotting as indicated. Data are representative of three independent experiments. Histogram indicates the relative level of PEAK3 p-S69 as mean ± S.E.M. over three independent experiments. ∗ p < 0.05, ∗∗ p < 0.01, by ratio paired t test.

Article Snippet: The following reagents/inhibitors were used in this study: lambda protein phosphatase (New England Biolabs, P0753S), animal-free recombinant human EGF (PeproTech, AF-100–15), insulin from bovine pancreas (Merck, I-1882), CaMKII inhibitor RA306 (custom synthesized by Reagency, RGNCY-0117, 1 μM final), PKA Inhibitor 14 to 22 Amide, Cell-Permeable, Myristoylated (Calbiochem, 476,485, 10 μM final), DMSO (Sigma, d8418, 0.1% final), and MedChemExpress-sourced Akt inhibitor MK-2206 dihydrochloride (HY-10358, 100 nM final) and trametinib (HY-10999, 10 nM final).

Techniques: Phospho-proteomics, Western Blot, Control, Inhibition, Derivative Assay, Knockdown

Journal: Materials Today Bio

Article Title: A bioinspired anisotropic anti-inflammatory scaffold enhances spinal nerve regeneration and neural circuit reconstruction via FGF13/Ca 2+ /CaMK2A/CREB pathway

doi: 10.1016/j.mtbio.2026.102929

Figure Lengend Snippet: Bioinformatics analysis and validation of the impact of A-NAC-AgNW-GM on neuroregeneration and neural circuit formation. (A) Volcano plot comparing gene expression in the A-NAC-AgNW-GM group to that in the SCl group, where the x-axis represents log2 (fold change) and the y-axis represents -log10 (P-value). (B) Partial GO analysis of the differentially expressed genes (DEGs). The x- and y-axes represent the number of genes and GO terms. (C) The circular plot displays the enrichment of differentially expressed genes in neuron-related functional categories and their expression changes (log2FC). (D) Bubble charts of the KEGG enrichment analysis for DEGs. The x and y axes represent the enrichment ratio and KEGG terms, respectively. (E) GSEA analyses were conducted on the calcium signaling pathway. (F) Heatmap showing DEGs within the calcium signaling pathway. (G) Western blot analysis of FGF13, CaMK2A, p-CaMK2A, CREB, and p-CREB protein expression in each group. (H-J) Statistical analysis of FGF13, p-CaMK2A/CaMK2A, and p-CREB/CREB protein intensity.

Article Snippet: Key antibodies used: iNOS (1:500, Affinity, AF0199), Arg-1 (1:500, Affinity, DF6657), CD68 (1:500, Santa Cruz, sc-20060), Iba-1 (1:1000, Abcam, ab178846), GFAP (1:1000, Abcam, ab4674), MAP-2 (1:500, Affinity, AF4081), MBP (1:500, Affinity, AF4085), 5-HT (1:500, Sigma, S5545), TH (1:1000, Abcam, ab112), Neun (1:1000, Abcam, ab104224), PSD95 (1:1000, Abcam, ab238135), Syn (1:500, Affinity, BF0348), CaMK2A (1:500, Cell Signaling, 50049), Cav1.2(1:500,proteintech, 21774-1-AP).

Techniques: Biomarker Discovery, Gene Expression, Functional Assay, Expressing, Western Blot