Fig. 3

Programmed knockout of BgAIF in Bge cells caused reduced adherence to primary sporocysts. a Representative micrographs of primary sporocysts co-cultured with Bge cells in our laboratory to profile the semi-quantitative scoring of the cell adhesion index (CAI): 1, no cells adhering to the surface of the sporocyst; 2, ≤ 10 cells adhering to the sporocyst; 3, > 10 cells < half of the sporocyst surface covered by cells or clumps of cells; 4, > half the sporocyst surface covered by Bge cells. b Representative micrographs indicate the reduced levels of ∆BgAIF-Bge cells adherence (right panel) in comparison to control, mock-transfected Bge cells (left panel) to the co-cultured sporocysts. c Bar chart to present the CAI values from control (mock-transfected) ∆BgAIF-Bge cells during co-culture with primary sporocysts at a co-culture ratio of one sporocyst to 100 Bge cells; CAI = 2.66 ± 0.10 (mean ± SD; 476 sporocysts in total scored) for the mock-transfected Bge and 2.31 ± 0.23 for the ∆BgAIF-Bge cells (424 sporocysts in total scored); P = 0.0033, unpaired Student’s t-test; n = 7 biological replicates. d The cell adhesion score ranging from 1 to 4 of individual sporocysts. Cell adhesion scores 1, 2, 3 and 4 from mock cells were 2.5 ± 1.58 (SE), 29.28 ± 4.06, 20.85 ± 2.15 and 13.14 ± 2.55, respectively, whereas in the ∆BgAIF-Bge cells cell adhesion scores 1, 2, 3 and 4 were 8.86 ± 1.57, 31.42 ± 4.07, 13.28 ± 2.51 and 13.13 ± 2.55, respectively. There was a higher amount of sporocyst from the BgAIF edited-mixed cell population scored as ‘1′ than control cells with statistic significant by t-test (t ratio = 3.98, P = 0.001). Also, there was a lower amount of sporocysts scored as ‘3′ in ∆BgAIF-Bge cells compared with the control cells (t ratio = 2.40, P = 0.004)