Paraffin blocks were cut at 3 μm, sections were mounted and dried on Superfrost Plus slides (Thermo Scientific) avoiding temperatures above 37°C. After dewaxing and rehydration, sections were incubated in one of the HIER buffers at different temperatures [20 min at 96°C in a steam cooker (Braun) or 90 min at lower temperatures in a water bath, details in Table 1].