Quant centre software

Immunofluorescence labelling of CD68 and CCR2 was performed on 2 µm cryo slides, labelling of NFκBp65 and α-amylase was performed on 2 µm paraffin slides. The primary antibodies were used in a 1:100 dilution and incubated over night at 4 °C. The secondary antibodies were used in a 1:200 dilution, for 1 h at room temperature. Nuclear staining was performed by DAPI. Slides were scanned by slide scanner (Pannoramic MIDI BF/FL, Sysmex Norderstedt, Germany). Quantification of positive cells was performed by Quant centre software from Sysmex, positive cells were calculated against total cell number.

Flow cytometry was analysed by FlowJo Software (BD Biosciences). All graphs were prepared by GraphPad Prism and SigmaPlot. Histology was evaluated by Quant centre software from Sysmex. Bubble chart was prepared by R Studio.

Quantification of apoptotic cells was performed by TUNEL assay (FragEL™ DNA fragmentation detection kit from Millipore, catalog no. QIA39–1EA) using paraffin sections. Slides were scanned with slide scanner (Pannoramic MIDI BF/FL, Sysmex Norderstedt, Germany). Quantification of apoptotic cells was performed by Quant centre software from Sysmex, TUNEL positive cells were calculated against total cell number^{37 (link)}.