Fig. 4
Panobinostat impacts on cellular differentiation via EGFR. A After treatment of cells with panobinostat (10xIC50: 100 nM for A204, and 250 nM for VAESBJ, GRU1) for 8 and 24 h, changes in mRNA expression levels for EGFR, FGFR1 and FGFR2 were determined in N ≥ 3 biological replicates in 3 technical replicates, normalized on GAPDH expression (top panel; graph represents means ± SEM; non-parametric Kruskal-Wallis test, p < 0.0001 for EGFR, FGFR1 and p = 0.0001 for FGFR2). Similarly, expression changes of FGFR2 3b and -3c mRNA isoforms were determined (bottom panel; graph represents means ± SEM; non-parametric Kruskal-Wallis test, p = 0.0024 (FGFR2 3b), p = 0.0031 (FGFR2 3c)). B Protein expression of EGFR after panobinostat treatment for 24, 48 and 72 h. C Cells were maintained in MEM and treated with panobinostat at 10/100 nM for A204, and 25/250 nM for VAESBJ and GRU1 (IC50/10xIC50) for 24 h, and stimulated with EGF to determine expression of EMT-related proteins by Western blot. D Transcript expression changes of EMT transcription factors SNAIL, SLUG and ZEB1 were measured after treatment of cells with panobinostat (10xIC50: 100 nM for A204, and 250 nM for VAESBJ, GRU1) for 8 and 24 h in N = 3 biological replicates in 3 technical replicates, normalized on GAPDH expression (graph represents means ± SEM; non-parametric Kruskal-Wallis test, p = 0.0029 (SNAIL), p = 0.01 (SLUG) and p = 0.0048 (ZEB1)). E Serum-starved cells were treated with panobinostat at above indicated respective IC50 and 10xIC50 doses for 24 h, in the presence or without EGF, to confirm SNAIL upregulation on the protein level by Western blot