Fig. 4: SET8 inhibition reduces cisplatin (CP)-induced apoptosis, along with preservation of PTEN in TKPTs.

TKPTs were treated with UNC0379 for 1 h (A–H) or transfected with siRNA targeting SET8 (siSET8) or control siRNA (siCON) (I-M) for 24 h and then exposed to cisplatin for 24 h. B Cells were conducted for TUNEL staining (A), and TUNEL (+) cells are shown. DAPI staining was used to localize nuclei in TKPTs. Scale bars = 50 μm. C Cell viability was detected after 24 h by cell counting kit 8 (CCK8). Cell lysates were prepared and subjected to immunoblot analysis with antibodies as indicated (D, I). The expression levels of all the proteins were quantified by densitometry. Cleaved cas3 (E), PTEN (F), and SET8 (G) were normalized by GAPDH; H4K20me1 (H) was normalized by H4; Cleaved cas3 (J), PTEN (K), and SET8 (L) were normalized by Tubulin; H4K20me1 (M) was normalized by H4. Cells were treated as described above. The extracted RNA cell lysates were subjected to quantitative RT-PCR using the primers listed in Supplementary Table 3. mRNA expression levels of SET and PCNA were normalized relative to mouse β-Actin and analyzed using the 2^−ΔΔCt method (N–Q). Data are represented as the mean ± SEM of at least three experiments. *P < 0.05, **P < 0.01, ***P < 0.001.