Fig. 2
Expression of Lr-pgrn genes in normal lamprey tissue after pathogen stimulation. A The expression of Lr-pgrn genes in the brain, kidney, heart, and other lamprey tissues was measured with qRT-PCR. B The distribution of Lr-PGRN-S1 in the gill, supraneural body, kidney, and intestine tissues was measured by immunohistochemistry. The two expressions were observed in a ×10 and ×40 magnification field of view, respectively. Normal rabbit IgG was used as negative control. C Lr-PGRN-S1 protein expression in lamprey cells by FACS analysis. Representative contour plots were gated on Lr-PGRN-S1-positive cells (green line). The black line indicates cells with the IgG isotype control. D Immunofluorescence detection of the cellular location of Lr-PGRN-S1. Light blue: nucleus; light green and red: Lr-PGRN-S1. E The expression level of Lr-pgrn genes in lampreys after S. aureus and V. anguillarum stimulation during 0, 4, 12, 24, and 48 h as determined by qRT-PCR and normalized to gapdh expression. F Interaction of rLr-PGRN-S1 with rL-TNFR as determined by BIAcore analysis. The KD values were measured using the BIA evaluation version 3.0 software. The data are presented as mean ± standard deviation (SD) based on three independent samples with three replicates per sample. Probabilities of statistical differences between experimental groups in all bar graphs were determined by Student’s t-test. *P < 0.05, **P < 0.001, ***P < 0.0001