Figure 4.

LV and AAV Testing in BALMs

(A) AAV and LV vectors. rAAV-LK03-CMV-eGFP (left) and LNT-LXR-Nluc/eGFP and LXR with its agonist T0901317 (right).

(B) Bioluminescence imaging of BALM1 transduced with LNT-LXR-Nluc/eGFP and agonist activation show luciferase expression after PV perfusion with a luciferase substrate in the DECs seeded lobes at day 25 (1 min exposure).

(C) Bioluminescence quantification of BALM1-3 after luciferase substrate perfusion (1 min exposure) at day 25.

(D) Immunostaining of GFP shows positive hiHEPs when transduced with LNT-LXR-Nluc/eGFP (BALM2, top panel) or with rAAV-LK3-CMV-eGFP (BALM4, bottom panel). Higher magnification images of merged channels in the right panel correspond to the delineated lower magnification images. Nuclei stained with DAPI. Scale bar, 50 μm.

(E) Bioluminescence fold change of secreted nanoluciferase in the media of BALM1-3 shows luciferase secretion upon LXR induction by its agonist at days 25–26. Untransduced negative control BALM (BALM0).

(F) Bioluminescence fold change of secreted nanoluciferase in the media of PHHs transduced with LNT-LXR-Nluc/eGFP for 24 h followed by 48 h incubation with 1 μM agonist shows luciferase secretion in PHHs.

See also Figures S4 and S5 and Tables S2 and S3.