Fig. 4: MYC is lost rapidly upon the inhibition of HDAC10.

A Pathway analysis of RNA sequencing data of RS4-11 cells treated with 15 µM PZ48 for 8 h (see Fig. 2A) using the Enrichr database. Shown are the MSigDB Hallmark 2020 data set (upper) and the NCI Nature 2016 data set (lower). Genes that we found downregulated in RNA sequencing analyses were used as input and an FDR < 0.05 was set; p-values increase from top to bottom. B Immunoblot analysis of RS4-11 cells that were treated with 15 µM PZ48 for increasing time periods (4 h, 8 h, 16 h, 24 h, n = 3). β-actin serves as loading control. C Quantification of B showing the mean ± SD values of 3 independent experiments. D RS4-11 cells were exposed to cycloheximide (CHX, left, 20 µM) and actinomycin D (ACTD, right, 2 µg/ml) for 2 h, 4 h, and 6 h. Immunoblot was done for POLD1 and MYC; GAPDH serves as loading control (n = 3). E Immunoblot was conducted with lysates from RS4-11 cells that were treated with increasing doses of MYCi361 (5 µM, 10 µM, 15 µM) in comparison to PZ48 (5 µM, 10 µM, 15 µM) for 24 h. GAPDH serves as loading control (n = 3). F RS4-11 cells were treated with MYCi361 (5 µM, 10 µM, 15 µM) for 24 h in comparison to untreated negative control cells (-) and treatment with 15 µM PZ48 (+) as positive control for apoptosis induction. Flow cytometry was used to detect Annexin-V/FITC and PI as early/late apoptosis markers (n = 3, mean+SD; two-way ANOVA; ****p ≤ 0.0001).