Figure 6: Validation of aberrant CA12 upregulation in FLC.
(a) CA12 RNA levels quantified by RNA-seq in non-tumor adjacent liver, primary FLC tumors, and metastatic FLC tumors in an independent cohort of FLC tumors17. (b) CA12 expression quantified by qRT-PCR in cultured spheroids from a FLC PDX model and Huh7, a human HCC cell line. CA12 expression was normalized to RPS9. Statistical significance determined by Student’s two-tailed t-test. Data shown are mean ± SD and display one representative experiment of biological replicates (n = 3 for Huh7, n = 6 for FLC PDX spheroids) of 4 total experiments. (c) CA12 expression measured by western blot in cultured spheroids from a FLC PDX model and Huh7. Uncropped blots are presented in Supplementary Fig. S14. (d) CA12 expression measured by immunohistochemistry in cultured spheroids from a FLC PDX model, xenograft tissue from a FLC PDX model, and normal human adult liver tissue. Magnification, 40x (top right), 20x (rest). (e) CA12 expression measured by immunohistochemistry in primary FLC and HCC. Magnification, 20x. (f) Table showing summary of samples with CA12 positive staining by IHC. NML, non-malignant liver. *p < 0.05, ****p < 0.0001 (Mann-Whitney U test).
