Journal: Molecular Cancer

Article Title: Panobinostat reduces hypoxia-induced cisplatin resistance of non-small cell lung carcinoma cells via HIF-1α destabilization

doi: 10.1186/1476-4598-14-4

Figure Lengend Snippet: Activation of apoptosis in NSCLC cells upon co-treatment with cisplatin and panobinostat. (A) H23 cells treated for 24 hours with single substances or with combination of both were stained with Hoechst 33342 in order to determine the level of chromatin fragmentation as an indicator of apoptosis. Cells were observed under the fluorescent microscope and representative photographs were made. Note markedly increased chromatin fragmentation in co-treated cells, those effects being better visible in boxed area at higher magnification. Percentage of fragmented nuclei in relation to total number of cells is shown (three independent experiments performed in triplicates). Scale bar = 50 μm. (B) Upon treatment H23 cells were stained with propidium iodide (PI) and analyzed by FACS for the sub-G1 peak characteristic for apoptosis. Data analysis was performed by ModFit LT 3.3 software package. Representative figures (left panel) and mean values (right panel) are shown. (C, D) NSCLC cells were treated with indicated concentrations of panobinostat, cisplatin or with combination of both, harvested in RIPA buffer and cell lysates were analyzed by immunoblotting using antibodies against different apoptosis markers. Upon stripping β-actin was used as loading control. P, panobinostat (16 nM); C, cisplatin (16 μM); P + C (16 nM panobinostat + 16 μM cisplatin); NOX, normoxia; HOX, hypoxia; Cl. PARP = cleaved PARP.

Article Snippet: For qRT-PCR 20 ng of each cDNA was used and reactions were performed by using following TagMan® assays on demand commercially available from Applied Biosystems: HDAC1, #Hs00606262_g1; HDAC2, #Hs00231032_m1; HDAC4, #Hs01041638_m1; HDAC5, #Hs00608366_m1; HDAC6, #Hs00195869_m1; HDAC8, #Hs00954353_g1; ß-actin, #Hs99999903_m1.

Techniques: Activation Assay, Staining, Microscopy, Software, Western Blot, Stripping Membranes, Control

Journal: Scientific reports

Article Title: The hydroxamate based HDAC inhibitor WMJ-J-09 induces colorectal cancer cell death by targeting tubulin and downregulating survivin.

doi: 10.1038/s41598-025-04714-w

Figure Lengend Snippet: Fig. 2. WMJ-J-09 disrupted microtubule assembly through HDAC inhibition. (A) Microtubule formation in HCT116 cells treated with colchicine, paclitaxel, WMJ-J-09, or vehicle, detected by confocal immunofluorescence analysis with β-tubulin-staining images in the upper panel, DAPI-staining images in the middle panel, and merged images in the bottom panel. (B) Immunoblotting result of polymerized tubulin in HCT116 cells treated with colchicine, paclitaxel, WMJ-J-09, or vehicle (C, D) Immunoblot result of α-tubulin acetylation induced by WMJ-J-09 in either a concentration-dependent (C) or time-dependent manner (D). (E) Immunoblot result of α-tubulin acetylation in WMJ-J-09-treated HCT116 cells with HDAC6 or HDAC8 overexpression. Each band intensity was quantified, and the total α-tubulin level normalized the fold changes of α-tubulin acetylation. Error bars, mean ± S.E.M. (shown only for independent replicate experiments with n ≥ 5).

Article Snippet: HDAC8 Flag was a gift from Eric Verdin (Addgene plasmid # 13825; http://n2t.net/addgene:13825; RRID:Addgene_13825) 64.

Techniques: Inhibition, Immunofluorescence, Staining, Western Blot, Concentration Assay, Over Expression