May gr nwald s eosine methylene blue solution

Morphological changes mediated by FX-9 exposure were analyzed through May-Grünwald-Giemsa staining. The four PCa cell lines as well as benign hTF-8 and 1801 cells were seeded in 60 cm² cell culture dishes (approx. 1 × 10⁶ cells per dish), which contained two microscope slides. After the cells reached about 80% confluence, the microscope slides were transferred to new dishes with FX-9 or DMSO for the control cells. Based on MTS assay PC-3, LNCaP and 0846 were incubated with 5.0 µM FX-9 and CT1258 with 2.5 µM FX-9 for 72 h. Benign cell lines hTF-8 and 1801 were also exposed to 5 µM FX-9 for 72 h. Microscope slides were air dried at room temperature. The slides were stained in May-Grünwald’s eosine-methylene blue solution (Merck, Darmstadt, Germany) for 200 s and rinsed with double distilled water. Afterwards, they were stained for 10 min in Giemsa’s azur eosin methylene blue solution (Merck, Darmstadt, Germany) and rinsed thoroughly with double distilled water again twice. The slides were left to air dry before analysis.

Mouse E-cadherin⁺;IgE⁺ cells were spun down at 800 rpm on an object glass using a Shandon Cytospin II (Thermo Scientific), air dried and fixed in methanol for 5 min. Samples were stained with May–Grünwalds eosine-methylene blue solution (Merck, Rahway, NJ, USA) for 10 min and Giemsa dilution for 20 min.

Morphological changes mediated by PDA were analyzed by May-Grünwald-Giemsa staining.
Microscope slides were placed in 60 cm² cell culture dishes and covered with 10 ml of culture medium. Per dish and per cell line, 1 x 10⁶ cells were seeded. On the following day, the microscope slides were transferred to new dishes with incubation medium or 0.15% DMSO for the control cells. The slides were exposed to the PDA agents in two different settings. For the first setting, the slides were incubated with 15 μM PDA-66 or PDA-377 for 24 h, an application equal to the cells used in the transcriptomic analyses. For the second setting, the slides were incubated with 5 μM PDA-66 or PDA-377 for 72 h, a concentration that showed moderate effects in all cell lines in live cell imaging. Microscope slides were washed with PBS and air dried at room temperature. The slides were stained in May-Grünwald's eosine-methylene blue solution (Merck KGaA, Darmstadt, Germany) (undiluted) for 5 min and rinsed with tap water. Afterwards, they were stained for 15 min in Giemsa's azur eosin methylene blue solution (Merck KGaA) (1:10 dilution) and rinsed thoroughly with tap water again. The slides were left to air dry before analysis.