Figure 1.
Creation of an inducible LucOS-expressing mouse HCC cell line
(A) Diagram showing how doxycycline induces the TetOn system to express luciferase and OVA MHC-I and -II antigens (LucOS). Blue arrows represent turning on of genes.
(B) Luciferase activity of the Hep55.1c cell line transduced with the Tet3G-LucOS expression system incubated with increasing concentrations of doxycycline.
(C) Intracellular IFNg production in OT-I CD8+ T cells co-incubated with Hep55.1c-Tet3G-LucOS cells as well as doxycycline and IFNg (10 ng/mL). NC means negative control and was Hep55.1c cells not transduced with the Tet3G-LucOS system.
(D) FACs plot showing OVA257-264 tetramer-positive CD8+ T cells from liver lymphocytes of mice treated with dox water or control water.
(E) Bar graph of (D)
(F and G) Representative picture and (G) tumor weight of mouse livers or tumors 10 days after an intrahepatic injection of Hep55.1c-Tet3G-LucOS given normal or doxycycline water. Low means low antigen expression due to no dox water while high means high antigen expression due to dox water.
(H) Correlation of tumor weight and percentage of tetramer-positive CD8+ T cells.
(I) Growth curves of flank Hep55.1c-Tet3G-LucOS tumors in mice given doxycycline water and depleted for CD4+ and/or CD8+ T cells (n = 5 per group). All pooled data presented as mean with SD. One-way ANOVA with Tukey correction for (B), (C) & (I) and unpaired t test for (E) & (G) ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001.