Journal: bioRxiv

Article Title: Mechanistic Insights into TYSM Protein Regulation by mTORC2 in Response to Chemotherapy

doi: 10.1101/2025.05.06.652447

Figure Lengend Snippet: (A) Lysates from MDA-MB-231 cells treated or not with 50 µM 5-FuDR or 50 nM Rapamycin for 24 hours were stained with the indicated antibodies. (B-C) Bar graphs showing the results of densitometric analysis of the protein levels under different conditions obtained in A. The data are presented as the means ±SEMs. ANOVA, multiple comparisons: Dunnett test. p-AKT1 (F (3, 8) = 4.06, P<0.05), p-P70SK (F (3, 8) = 17.5, P<0.001), TYMS (F (3, 8) = 16.34, P<0.001). (D) Lysates from MCF7 cells treated or not with 50µM 5-FuDR, 50nM Rapamycin for 24 hours were stained with the indicated antibodies. (E-F) Bar graphs showing the results of densitometric analysis of the protein levels under different conditions obtained in D. The data are presented as the means ±SEMs. ANOVA, multiple comparisons: Dunnett test. p-AKT1 (F (3, 8) = 18.5, P<0.0005), p-P70SK (F (3, 8) = 76.9, P<0.0001), TYMS (F (3, 8) = 124.1, P<0.0001).

Article Snippet: The following inhibitors were used: 5-fluorouracil (Sigma Aldrich, #F6627), 5-Fluoro-2’-deoxyuridine (Thermo Scientific Chemicals, #L16497.ME), 5-fluorouridine (Thermo Scientific Chemicals, #J62083.03), Rapamycin (Thermo Scientific Chemicals, #J62473.MC).

Techniques: Staining

Journal: Cell reports

Article Title: De novo recovery of Ghana virus, an African bat Henipavirus, reveals differential tropism and attenuated pathogenicity compared to Nipah virus

doi: 10.1016/j.celrep.2026.117074

Figure Lengend Snippet: (A–E) Dose-response curves are shown for various small-molecule inhibitors against rGhV M-GS* (purple) and rNiV (orange). CHO-B2 cells were infected at an MOI of 0.05 and treated with (A) EIDD-2749, (B) GHP-88309, (C) remdesivir, (D) EIDD-1931 (molnupiravir active form), or (E) favipiravir (T-705). (F and G) Additional dose-response curves were measured in primary human astrocytes for (F) EIDD-2749 and (G) GHP-88309 under the same MOI and assay conditions. (H) Summary of half-maximal inhibitory concentrations (IC 50 ) derived from the dose-response curves in (A–G). Each row indicates the respective compound, virus, calculated IC 50 , corresponding 95% confidence interval, and the cell type in which the assay was performed. IC 50 values were determined in GraphPad Prism by nonlinear regression of (inhibitor) vs. normalized response. All experiments were conducted in biological triplicate. Error bars depict standard deviation.

Article Snippet: EIDD-2749 (4′-Fluorouridine) , MedChemExpress , Cat# HY-146246.

Techniques: Infection, Derivative Assay, Virus, Standard Deviation

Journal: bioRxiv

Article Title: P-TEFb controls cell fate upon p53 activation by modulating intrinsic apoptosis pathway

doi: 10.1101/2022.05.29.493929

Figure Lengend Snippet: Synthetic lethality of P-TEFb inhibitor NVP-2 with MDM2 inhibitor Nutlin-3a and antimetabolites is p53-dependent. (A-D) 8 × 6 matrices with combinatorial titrations of NVP-2 (green) with Nutlin-3a (red) and 5-Fluorouracil (5-FU; blue) at indicated doses to test for the synthetic lethality of compounds in HCT116 TP53 +/+ and HCT116 TP53 cells, depicting cytotoxicity (top) and synergy (bottom) of the combinations. Cytotoxicity values obtained at 48 hr of the treatments using CellTox Green assay were normalized to the DMSO control and are presented as percentages of the maximum cytotoxicity which was set at 100 %. Results represent the average of independent experiments (n = 3). Combinations with the highest Bliss synergy scores in HCT116 TP53 +/+ cells are highlighted (gold). (E,F) Cytotoxicity of HCT116 TP53 +/+ and HCT116 TP53 -/- cells treated with the DMSO control (grey), NVP-2 (10 nM; green), Nutlin-3a (10 μM; red), and antimetabolites 5-Fluorouracil (5-FU; 50 μM), 5-fluorouridine (5-FUR; 0.5 μM) and 5-fluorodeoxyuridine (5-FUdR; 0.15 μM) (blue) alone and in combination (gold) as indicated for 48 hr measured using CellTox Green assay. Results are presented as fluorescence values relative to the DMSO control and plotted as the mean ± s.e.m. (n = 3). **, P < 0.01; n.s., non-significant, determined by Student’s t test. See also .

Article Snippet: 5-Fluorouridine (FUR), Floxuridine (5-FUdR) were from MCE Chemicals.

Techniques: CellTox Assay, Control, Fluorescence

Journal: bioRxiv

Article Title: P-TEFb controls cell fate upon p53 activation by modulating intrinsic apoptosis pathway

doi: 10.1101/2022.05.29.493929

Figure Lengend Snippet: Synthetic lethality of p53 activation and P-TEFb inhibition remains effectual in spheroid culture system. (A) Representative images of HCT116 TP53 +/+ spheroid cultures treated with DMSO, Nutlin-3a (10 μM), NVP-2 (10 nM), and 5-Fluorouracil (5-FU; 25 μM) alone and in the combinations as indicated. Spheroids were formed for 48 hr, exposed to the treatments for 96 hr and stained with a mixture Calcein AM (1 μM), DRAQ7 (6μM) and Hoechst (1x) prior to confocal microscopy imaging. Composite images of all three staining are shown on the right. Scale bar, 100 μm. (B-F) Apoptosis of the indicated HCT116 TP53 R248W/+ , HCT116 TP53 R248W/- and HCT116 TP53 +/+ cell spheroid cultures treated with DMSO (grey), Nutlin-3a (10 μM; red), NVP-2 (10 nM; green), 5-fluorouridine (5-FUR; 0.5 μM) and 5-fluorodeoxyuridine (5-FUdR; 0.15 μM), 5-Fluorouracil (5-FU; 25 μM) and DT-061 (10 μM; green) alone and in the indicated combinations (gold). Spheroids were formed for 48 hr prior to the treatments. Results obtained at the time points indicated below the graphs using RealTime-Glo Annexin V assay are presented as luminescence values relative to the DMSO control values at 2 hr and plotted as the mean ± s.e.m. (n = 3). *, P < 0.05; **, P < 0.01; ***, P < 0.001, determined by Student’s t test using Nutlin-3a and Nutlin-3a + NVP-2 (B), 5-FUR and 5-FUR + NVP-2 (C), 5-FdUR and 5-FdUR + NVP-2 (D), Nutlin-3a and Nutlin-3a + DT-061 (E), and 5-FU and 5-FU + DT-061 (F) data sets.

Article Snippet: 5-Fluorouridine (FUR), Floxuridine (5-FUdR) were from MCE Chemicals.

Techniques: Activation Assay, Inhibition, Staining, Confocal Microscopy, Imaging, Annexin V Assay, Control