Journal: Nature Communications

Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation

doi: 10.1038/s41467-024-53178-5

Figure Lengend Snippet: a , b The gels illustrate the resulting epichaperome formation and the phosphorylation status of HSP90 in lysed MDA-MB-468 epichaperome-positive cancer cells treated with CK2 inhibitors. Detection of epichaperome components was done through SDS–PAGE (total protein levels) and native PAGE followed by immunoblotting. a The effect of CX4945 treatment, while b depicts the effect of CIBG-300 treatment. Vehicle-treated cells serve as controls. CK2α levels are shown to verify that inhibitor treatment effects are independent of changes in CK2α expression levels. c Same as in ( a , b ) for CK2α knockdown using dose-dependent siRNAs in MDA-MB-468 cells. CK2α levels, knockdown efficiency control. d The indicated CK2 constructs were used to transfect HEK293 cells. CK2α, catalytic subunit; CK2β, regulatory subunit; kinase-dead mutant CK2 K68M α. HA tag and CK2α levels, transfection efficacy control. a – d Gel images are representative of three independent experiments. Source data are provided as a Source data file. e Schematic summary. CK2’s phosphorylation activity directly influences the stability and assembly of epichaperomes. These findings confirm the functional role of HSP90 phosphorylation at these specific serine residues in epichaperome formation and posit CK2 as a likely physiological candidate behind epichaperome formation.

Article Snippet: CX4945 (Silmitasertib) was purchased from MedChemExpress (Cat No. HY-50855), and CIBG-300 was obtained from Sigma-Aldrich (Cat No. SML3143).

Techniques: Phospho-proteomics, SDS Page, Clear Native PAGE, Western Blot, Expressing, Knockdown, Control, Construct, Mutagenesis, Transfection, Activity Assay, Functional Assay

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Rutaecarpine alleviates inflammation and fibrosis by targeting CK2α in diabetic nephropathy.

doi: 10.1016/j.biopha.2024.117499

Figure Lengend Snippet: Fig. 6. Prediction of molecular targets of Rut. (A) Profiling of the predicted protein targets of Rut via DS 2021. The y-axis represents Rut, and the x-axis indicates the predicted pharmacophore models (pharmacological targets) of Rut. The color from blue to red represents a gradual increase in fit values. (B) Molecular docking. (C) CETSA analysis of the stabilization of CK2α with or without Rut treatment. (D) SPR test using Biacore demonstrating the stable fit of the interaction between Rut and CK2α. (E) Immunohistochemistry analysis of CK2α of mouse kidney. Scale bars = 50μm. Data are presented as the mean ± SD, n=6. (F) Western blot analysis of CK2α protein levels of mouse kidney(a) (Data are presented as the mean ± SD, n=6) and SV40 cells(b) (Results are presented as the mean ± SD of three independent experiments).

Article Snippet: CK2 inhibitor CX4945 (10 μM final concentration; Selleck Chemicals, Cat#S2248).

Techniques: Immunohistochemistry, Western Blot

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Rutaecarpine alleviates inflammation and fibrosis by targeting CK2α in diabetic nephropathy.

doi: 10.1016/j.biopha.2024.117499

Figure Lengend Snippet: Fig. 7. Rut failed to suppress HG-induced ECM accumulation and activation of the TGF-β1/ Smad3 signaling pathway and attenuate HG-induced inflammation and NF-κB activation in CK2α-silenced SV40 MES-13 cells. (A) Western blot analysis of CK2α levels in SV40 cells. (B) Real-time PCR analysis of CK2α mRNA expression in SV40 cells. (C) Western blot analysis of FN, Col-IV, α-SMA, TGF-β1, p-Smad3, and Smad3 protein levels in SV40 cells. (D) Real-time PCR analysis of IL-1β, TNF-α, and MCP-1 mRNA expression in SV40 cells. (E) Western blot analysis of p-NF-κB p65, NF-κB p65 protein levels of SV40 cells. Results are presented as the mean ± SD of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: CK2 inhibitor CX4945 (10 μM final concentration; Selleck Chemicals, Cat#S2248).

Techniques: Activation Assay, Western Blot, Real-time Polymerase Chain Reaction, Expressing

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Rutaecarpine alleviates inflammation and fibrosis by targeting CK2α in diabetic nephropathy.

doi: 10.1016/j.biopha.2024.117499

Figure Lengend Snippet: Fig. 8. Rut exerted better protective effects against HG-induced cell injury than the classical CK2α inhibitor CX4945. (A-a) Effect of varying concentrations of CX4945 on SV40 cell viability (MTT assay). (A-b) CX4945 inhibits the proliferation in HG-treated SV40 cells (MTT assay). (B) Western blot analysis of FN, Col-IV, and α-SMA protein levels in SV40 cells. (C) Western blot analysis of TGF-β1, p-Smad3, and Smad3 protein levels in SV40 cells. (D) Real-time PCR analysis of IL-1β, TNF-α, and MCP-1 mRNA expression in SV40 cells. (E) Western blot analysis of p-NF-κB p65, NF-κB p65 protein levels of SV40 cells. Data represent the mean ± SD for three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: CK2 inhibitor CX4945 (10 μM final concentration; Selleck Chemicals, Cat#S2248).

Techniques: MTT Assay, Western Blot, Real-time Polymerase Chain Reaction, Expressing