Samples were run for 20 h at 20 V and stained afterwards with ethidium bromide (0.5 μg ml−1).
PCR product was run on a 10% acrylamide gel (BioRad) and stained afterwards for 10 min in a 0.5μg/ml ethidium bromide (Sigma-Aldrich) solution.
In parallel, 20 μl of each eluate was analyzed by 7% Tris-Acetate SDS-PAGE gels (Life Technologies, Grand Island, NY, USA) and stained afterwards by using EZBlue Gel Staining Reagent (Sigma, St . Louis MO, USA).
To control for the accuracy of the puncture the sections were stained (cresyl violet) afterwards.
Protein samples in SDS-sample buffer were boiled and loaded (10 μg/lane) on a 12%% polyacrylamide gel and afterwards stained using colloidal Coomassie (Sigma Aldrich, Germany).
The organ was first immersed in saline solution, then fixed in methanol for 3 min, and afterwards stained with the Romanovsky method (Giemsa stain-Merck,Germany) used routinely in the lab [ 25].
To verify that the PI uptake is not due to cell death, the cells were stained with Trypan blue afterwards and blue stained cells were excluded from calculations.
Afterwards the sections were coated with DAB staining reagent for 10 min at room temperature.
Afterwards, the plates were washed rigorously with ddH2O and staining images were collected under a scanner (Hewlett-Packard Scanjet 2400; Hewlett-Packard, Palo Alto, CA, USA).
FITC-labelled HRP-conjugated anti-rabbit IgG (Sigma, Dorset, UK) was added afterwards, followed by mounting using 6-diamidino-2-phenylindole (DAPI; staining nuclei in blue) containing anti-fade ProLong Gold reagent (Life Technologies Ltd ,Paisley, UK).
