Fig 2. Immune cell activation after stimulation with M. luminyensis.
Cytokine release after stimulation of 1×105 PBMCs (A) as well as 1×105 moDCs (B) with 1×106 and 1×107 M. luminyensis or M. stadtmanae cells for 20 h was quantified using commercial ELISA-Kits. Unstimulated cells (Med. ctrl.) were used as negative controls. Depicted data are means of at least 3 independent biological replicates with their respective standard errors of the mean (SEM). Values are compared to medium control. ns: not significant, * P≤0.05, ** P≤0.01, *** P≤0.001 (one-way ANOVA with Bonferroni post hoc test). C) 1×105 moDCs were stimulated with 1×107 FITC-labeled methanoarchaeal cells in VI channel μ-slides for a period of 16 h. After incubation, moDCs were washed, fixed with 3% paraformaldehyde and DNA was labeled with Hoechst 33342. Images were captured using Leica SP5 confocal microscope with Leica confocal software and are representative of the respective samples (three independent biological replicates). D) Phagocytosis rate of M. stadtmanae and M. luminysensis by moDCs was determined by counting phagocytosed archaeal cells in image sections of three biological replicates (mean of counted moDCs: Medium ctrl = 34, moDCs stimulated with M. stadtmane = 56, moDCs stimulated with M. luminyensis = 45). Values are compared to each-other. *** P≤0.001 (two-tailed unpaired t-test).