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: The MDA-MB-468 (female, breast cancer cell line, HTB-132, RRID: CVCL_0419), ASPC1 (female, pancreatic cancer cell line, CRL-1682, RRID: CVCL_0152), NCI-H1975 (female, non-small cell lung cancer cell line, CRL-5908, RRID: CVCL_1511), Daudi (male, B lymphoblast cell line, CCL-213, RRID: CVCL_0008), MRC5 (male, lung fibroblast cell line, CCL-171, RRID:CVCL_0440), CCD-18Co (female, colon fibroblast cell line, CRL-1459, RRID: CVCL_2379) and the Human Embryonic Kidney 293 (HEK293) cell line (CRL-1573, RRID: CVCL_0045), of female origin as determined by sequencing, were purchased from ATCC.
Techniques: SDS Page, Clear Native PAGE, Western Blot, Expressing, Knockdown, Control, Construct, Mutagenesis, Transfection, Activity Assay, Functional Assay