Fig. 2

AChE inhibition suppresses PCC proliferation and invasion in vitro. a-h T3M-4 and SU86.86 human pancreatic cancer cells (PCCs) were treated for 24 h, 48 h and 72 h with indirect parasympathomimetic drugs (physostigmine, pyridostigmine), with the direct parasympathomimetic carbachol or with acetylcholine (ACh) and analyzed for their viability via MTT assay, Graphs shows cell growth of human PCCs over time and area under curve (AUC) values for different treatment regimens (T3M-4: physostigmine/Physo 10 ng: *p = 0.0153; pyridostigmine/Pyrido 300 ng/100 μl:*p = 0.0175; for ACh: T3M-4 and SU86.86: 500 μM ***p < 0.0001; 1000 μM ***p < 0.0001; T3M-4-carbachol: 1 μM: **p = 0.0011, 10 μM: ***p < 0.0001, 100 μM: ***p < 0.0001, 1 mM: **p < 0.0022; SU86.86-carbachol: ***p = 0.0011, unpaired t-test of area under the curve/AUC). i AChE activity assay with the SU86.86 and T3M4 cell lines. j-l Representative photomicrographs of transwell chamber membranes with CFSE–stained SU86.86 PCCs after treatment with physostigmine, pyridostigmine or ACh. Graphs shows the percentage of treated migrated cells compared to solvent-treated treated controls (Physostigmine. 30 ng/μl *p = 0.0102 by unpaired t-test; pyridostigmine: 10 ng/μl *p = 0.0162; 30 ng/μl ****p < 0.0001; ACh: ****p  = 0.0006; by unpaired t-test). All experiments were performed in biological triplicates