Journal: Molecular cancer therapeutics

Article Title: Oncogenic cells of renal embryonic lineage sensitive to the small molecule inhibitor QC6352 display depletion of KDM4 levels and disruption of ribosome biogenesis

doi: 10.1158/1535-7163.MCT-23-0312

Figure Lengend Snippet: (A) PrestoBlue assay dose response curves display the sensitivity (IC50) of Wilms tumor cell lines (WiT49, 17.94, COGW408, and PDM182) and the tumor-forming embryonic kidney cell line, HEK293 to the KDM4 inhibitor QC6352. (B) Crystal violet assay demonstrates a dose-dependent response to QC6352 in WiT49 and HEK293. (C) Live-cell microscopy proliferation assay demonstrates reduced proliferation in QC6352-treated WiT49 and HEK293. (D) Scratch wound cell migration assay performed using phase-contrast live-cell microscopy demonstrates decreased migration in QC6352-treated WiT49 and HEK293. (E) QC6352 significantly reduces spheroid growth in WiT49 and HEK293. For C-E, the rightmost asterix indicates the timepoint at which separation in curves is statistically significant (p<0.05; paired two-tailed t-test) and persists throughout remaining indicated duration. (F) Tumor photographs and volume curves from HEK293 subcutaneous xenografts in NSG mice demonstrate a volumetric response to QC6352 (25 mg/kg oral gavage for three weeks) compared to vehicle control (paired two tailed t-test values shown).

Article Snippet: The favorable histology WT cell line PDM182 (HCM-BROD-0051-C64; ATCCPDM-182) was obtained from ATCC and cultured in Conditional Medium (Propagenix, 256–100).

Techniques: Prestoblue Assay, Wilms Tumor Assay, Crystal Violet Assay, Microscopy, Proliferation Assay, Cell Migration Assay, Migration, Two Tailed Test, Control

Journal: Cardiovascular Research

Article Title: Cyclophilin D-mediated regulation of the permeability transition pore is altered in mice lacking the mitochondrial calcium uniporter

doi: 10.1093/cvr/cvy218

Figure Lengend Snippet: Phosphorylation of S42-CypD decreases CRC and increases CypD association with ATP synthase and cell death. (A) CypD levels were measured between different stable cell lines containing S42A or S42D mutations. ATP synthase subunit α was used as a loading control. (B) Representative calcium uptake assay in CypD-KO MEFs infected with lentivirus containing mutated CypD, either phospho-resistant S42A or phosphomimic S42D. (C) Mean CRC values, measured as nmol of calcium per mg of protein, indicated that phosphorylation of CypD at S42 decreased CRC. Data are represented as mean ± SEM. *P = 0.005 (two-way ANOVA), n = 5 independent experiments. (D) Immunoprecipitation of ATP synthase pulled down more CypD in the S42D mutant than S42A. Data are represented as mean ± SEM. *P = 0.02 (Student’s t-test), n = 5 independent experiments. (E) Oxidative stress induced with 1 mM H2O2 caused more cell death in the phosphomimic S42D mutant than in the phospho-resistant S42A mutant. P < 0.001 (Two-way ANOVA), *indicates within time point, and †indicates overall effect of cell line; n = 6–7 independent experiments.

Article Snippet: CypD WT and S42A and S42D mutants were purchased from GenScript and sub-cloned into the lentiviral vector PLVX-puro (Clontech, Palo Alto, CA, USA).

Techniques: Phospho-proteomics, Stable Transfection, Control, Infection, Immunoprecipitation, Mutagenesis