Fig. 1: A pro-survival role of LATS1 in HCC cells in response to sorafenib treatment.

a Huh7 cells were transfected with indicated siControl and treated with DMSO vehicle or sorafenib (Srf) for 72 h. Cell death was analyzed by immunoblotting with indicated antibodies. Results represent three independent experiments. b Colony formation was determined in Huh7 cells expressing either shControl (shRNA targeting LacZ) or a shRNA against LATS1 and exposed to sorafenib. Results represent three independent experiments. c Huh7 cells either expressing shControlor sh LATS1 were implanted into the flank of NSG mice. The mice were treated with placebo or sorafenib and tumor volumes were measured. n = 7–9 for each treatment cohort. Statistical significance was calculated using the R package compareGrowthCurves. d Loss of LATS1 expression results in impaired viability of HCC organoid lines in response to sorafenib (Srf). Results were pooled from three to five independent experiments. Statistical significance was calculated using two-tailed, paired t-test. e Schematic illustration of establishment of sorafenib-resistant cells by (1) step-wise increasing sorafenib concentration (Huh7-IR cells) and (2) consistent high concentration (Huh7-CR cells). f Analysis of LATS1 protein expression in sorafenib-resistant cells. Parental (Huh7p) and IR/CR cells were treated with Sorafenib (5 μM) for24 h. Cells were analyzed by immunoblotting with indicated antibodies. Results represent three independent experiments. g siRNA-mediated loss of LATS1 expression results in impaired colony formation of sorafenib-resistant Huh7 cells in response to sorafenib (Srf). Results represent three independent experiments. h, i The expression of LATS1 (h) and LATS2 (i) was determined in the TCGA database. Expression values were log2-transformed. Statistical analysis was calculated using two-tailed, unpaired Welch’s t-test. j, k Kaplan–Meier analysis of the TCGA database for the expression of LATS1. Overall survival (OS) (j) and disease-free survival (DFS) (k) of high and low patients are shown. Statistical significance was calculated using log-rank (Mantel–Cox) test. l LATS1 expression was determined by RNA sequencing of needle biopsies from patients before sorafenib treatment and during sorafenib treatment. mRNA levels of tumor biopsies were normalized to corresponding adjacent non-neoplastic tissue for each individual patient. Each dot represents one patient (Pre-sorafenib: n = 3 for responders and n = 6 for non-responders; On-sorafenib: n = 4 for responders and n = 6 for non-responders). Data represents log2 fold changes. Statistical significance was determined by Komogorov–Smirnov t-test.