Fig. 5 | Scientific Reports

Fig. 5

From: Integration of biomimetic organoid-on-chip and 2D models advances the mechanistic Understanding of STEAP3-mediated regulation in intestinal viral infection

Fig. 5

Colon organoids-on-chips for high-throughput screening of STEAP3’s inhibitory effects on viral infectivity. (A) Schematic showing the dimensions of the 3D-printed chip. (B) Top view of an individual well in the chip. (C) A graphic illustrating the actual chip. (D) Experimental procedures for constructing a biomimetic model of virus-infected colon organoids in the chip. (E) Detailed protocol for evaluating the effects of STEAP3 on viral infection in colon organoids-on-chip. Colon organoids were seeded in the chip and transfected with siControl or siSTEAP3 siRNA for 24 h. The siRNA-transfected organoids were then infected with the SARS-CoV-2 S entry virus for another 24 h. Total protein lysates were extracted from the organoids and subjected to the luciferase activity assay. (F) The expressional levels of STEAP3 in control or STEAP3 knockdown colon organoids. The uncropped and unprocessed blots are provided in Supplementary Fig. S19. (G-J) Colon cancer organoids were infected with a SARS-CoV-2 S entry virus encoding a spike protein in its wild-type form or with mutations at amino acid 614 (G614D), eight mutations (B.1.1.7), or two deletions (V501Y.V2) for 24 h. Luciferase activity was measured and quantified as the ratio of luciferase activity to protein concentration (n = 3). *p < 0.05 versus siControl-transfected cells using the student’s t-test. All data are shown as the mean (± s.d.).

Back to article page