Journal: Acta Dermato-Venereologica

Article Title: Possible Clinical Effects of Ketoconazole on Sorafenib-induced Hand–Foot Skin Reaction and Cytoprotection Mechanisms of Antifungal Agents against Multikinase Inhibitor-induced Keratinocyte Toxicity

doi: 10.2340/actadv.v105.40697

Figure Lengend Snippet: Detection of apoptosis-related factors in NHEK cell lysates following sorafenib and antifungal drug treatment. Detection of (A) cIAP-1 (70 kDa) and (C) cleaved caspase-3 (19 kDa) in the cell lysate was performed using western blotting. (B) Bands were semi-quantified using ImageJ, and data were expressed as the relative ratio of cIAP-1/β-actin. Statistical analysis was conducted using one-way ANOVA followed by Tukey’s test; ns indicates not significant. Sora or S: sorafenib, Keto: ketoconazole, Itra: itraconazole. All results are presented as the mean ± SD of triplicate experiments. (D) Various concentrations of sorafenib, with or without 50 μmM ZVAD or ZDEVD, were added to cultured NHEK cells. DMSO was used as the solvent for both sorafenib and the inhibitors. After overnight incubation, cell viability was assessed using the cell-counting kit-8 assay. Statistical analysis was performed using one-way ANOVA followed by Dunnett’s test; ** p < 0.01 and * p < 0.05 (vs DMSO (-)).

Article Snippet: Z-DEVD-FMK (ZDEVD) and Z-VAD-FMK (ZVAD) were from Selleck Chemicals (Houston, TX, USA).

Techniques: Western Blot, Cell Culture, Solvent, Incubation, Cell Counting

Journal: Cell Death & Disease

Article Title: Targeting POLRMT by IMT1 inhibits colorectal cancer cell growth

doi: 10.1038/s41419-024-07023-8

Figure Lengend Snippet: The primary human colon cancer cells, pCan1, underwent treatment with IMT1 (1 μM). Subsequently, they were cultivated for the designated duration, and the assessments including the Caspase-3 activity ( A ) and the Caspase-9 activity ( B ), expression of apoptosis-related proteins ( C ), and cytosol cytochrome C contents (ELISA assays, D ) were performed. Cell apoptosis was tested through nuclear TUNEL staining assays ( E ). Preceding IMT1 (1 μM) treatment for the specified period, pCan1 cells underwent pretreatment with zDEVD-fmk (50 μM) or zVAD-fmk (50 μM) for 1 h. Subsequent assessments included the evaluation of apoptosis via nuclear TUNEL staining assays ( F ) and the determination of cell death through Trypan blue staining ( G ). Other primary colon cancer cells (“pCan-2/pCan-3”) or immortalized colon cancer cells (HCT116) were exposed to IMT1 (1 μM) for a designated duration and assessments included the Caspase-3 activity ( H ) and nuclear TUNEL staining ( I ). “Veh” represents the vehicle control (0.1% DMSO). The data were presented as mean ± standard deviation (SD, n = 5), with *indicating P < 0.05 compared to the “Veh” treatment ( A – I ) and # indicating P < 0.05 compared to the IMT1 only treatment ( F , G ). The experiments were repeated five times, yielding consistent results. The scale bar is set at 100 μm.

Article Snippet: The pan-caspase inhibitor (zVAD-fmk) and the specific caspase-9 inhibitor zDEVD-fmk were procured from Calbiochem (Darmstadt, Germany).

Techniques: Activity Assay, Expressing, Enzyme-linked Immunosorbent Assay, TUNEL Assay, Staining, Control, Standard Deviation