Fig 2: ETO regulates SAMHD1 phosphorylation through the p53, p21 pathway MDM were treated with increasing concentrations of ETO and CTH. Cells were stained for MCM2 expression, acquired and analysed using the automated cell-imaging system Hermes WiScan and ImageJ. On average, 104 cells were acquired (n = 3, mean ± s.e.m.).MDM were treated with 5 µM ETO, lysed and immunoblotting performed to detect cell cycle/cell cycle arrest and DNA damage-associated proteins.Quantification of specific proteins band intensities from immunoblot in panel (B) using a CCD camera. Intensities of protein bands were normalized to intensity of actin protein band.MDM were treated with 5 µM ETO or 0.01 µM CTH for 18 h, lysed and immunoblotting performed to detect cell cycle/cell cycle arrest and DNA damage-associated proteins.Data information: (B–D) Each panel shows a representative example of three independent experiments.Source data are available online for this figure.

Fig 3: Western blot analysis for the target proteins. (A) The Western blot gels for p-p53, SMAD2/3, and SMA compared to ß-ACTIN. (B,C) Column charts for the p-p53 and SMAD2/3 in the experimental groups. Data are mean ± SD. †: versus the vehicle group, ‡: versus the LEFN 2.5 mg/kg group, and S: versus the LEFN 5 mg/kg group at p < 0.05.

Fig 4: Immunohistochemistry for p53 in the kidney specimens. (A) Images from the vehicle group show focal weak p53 staining, localized to periglomerular cells (in left panel) and focal minimal staining in tubular cells (right panel). (B) Images from the LEFN 2.5 mg/kg group show a very faint focal staining of p53 in glomerular areas (left panel) and weak staining in peritubular areas (arrow in right panel). (C) The LEFN 5 mg/kg group showed focal moderate or weaker staining of p53 encircling periglomerular areas (left panel), and this is also seen in peritubular and perivascular areas (right panel). (D) The LEFN 10 mg/kg group revealed moderate to strong staining of p53, with most staining in periglomerular tubules (left panel) and in areas surrounded by inflammatory cells infiltrate (right panel), arrows in all images indicate positive staining (E) Column chart for mean ± SD of the stained area % in kidney specimens from the experimental groups and data were analyzed using one-way ANOVA. †: versus the vehicle group, ‡: versus the LEFN 2.5 mg/kg group, and S: versus the LEFN 5 mg/kg group at p < 0.05.