Fig. 3

Genomic evidence of osmoregulation related genes in P. hakonensis and P. brandtii. A Genes and pathways involved in osmoregulation are shown in the colored circles and light blue box, respectively. PSGs are marked in pink circles, REGs are marked in purple circles. B Immunoblotting assay of prl in HEK293T cells. HEK293T cells were cultured in 6-well plates and transfected with indicated plasmids (3 μg each) for 24 h, and then the cells were harvested to perform immunoblotting. C prl-WT reduces cell crumbling and abscission after high osmotic pressure treatment. HEK293T cells were cultured in 6-well plates and transfected with indicated plasmids (3 μg each) for 8 h and then cultured under high osmolarity condition (600 mOsmol/kg) for 16 h and imaged under a microscope Nikon TE2000-U. D–E prl-WT reduces cell apoptosis in high osmolarity condition (600 mOsmol/kg) or low osmolarity condition (200 mOsmol/kg) detected by flow cytometry analysis (D), and statistical analyses were performed using the GraphPad Prism software (7.0) (E). HEK293T cells were cultured in 6-well plates and transfected with the indicated plasmids (3 μg each) for 8 h and then cultured under high osmolarity condition (600 mOsmol/kg) for 16 h (high group) or cultured under high osmolarity condition (600 mOsmol/kg) for 12 h followed by low osmolarity condition (200 mOsmol/kg) for 4 h (low group). Data show mean ± SD; Student’s two-tailed t test. *P < 0.05, **P < 0.01, ***P < 0.001, data from three independent experiments