Fig. 5

LIPH-LPA-LPAR axis enhanced PDAC glycolysis and viability by activating hypoxia and Hippo pathway. A KEGG pathway analysis showed enrichment of PI3K/AKT and Hippo pathway components in PANC-1 NC treated with PA (10 µM), compared to PANC-1 sh1-LIPH-transfected cells treated with PA (10 µM). B q-PCR showed that LIPH knockdown inhibited utilization of exogenous PA (10 µM) and downregulated CTGF expression at the mRNA level. C q-PCR showed that LIPH overexpression promoted utilization of exogenous PA (10 µM) as well as CTGF expression. D Immunoblot analysis of PDAC cells, with LIPH knockdown by sh1-LIPH or control, in the presence or absence of exogenous PA and LPA. E IF analysis of YAP1 localization of MIA PaCa-2 cells with LIPH overexpression by OE-LIPH vector and vector control in the presence or absence of FBS. F IF analysis of YAP1 and LIPH localization and expression level in PDAC patient tissues (red: YAP1, green: LIPH, blue: DAPI). G Immunoblot analysis of MIA PaCa-2 cells with LIPH overexpression by OE-LIPH or vector control. H Evaluation of the stability of YAP1 in MIA PaCa-2 cells stably transfected with OE-LIPH or vector control. Cells were treated with protein synthesis inhibitor cycloheximide (CHX; 50 μg/mL) and YAP1 abundance was examined by immunoblot of lysates of cells collected at the indicated times. I Abundance of YAP1 in cytosolic (Cyto) and nuclear (Nuc) fraction in MIA PaCa-2 cells stably transfected with OE-LIPH or vector control. Chip-qPCR analysis of abundance of YAP1 in HIF1A promoter in MIA PaCa-2 cells stably transfected with OE-LIPH or vector control. J Immunoblot analysis of MIA PaCa-2 cells with LIPH overexpression or vector control in the presence or absence of ki16425 (10 μM). K Subcutaneous tumor after the administration of ki16425 (10 mg/kg, qod) or corresponding solvent. Scale bar = 100 µm. *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001