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. Author manuscript; available in PMC: 2018 Apr 27.
Published in final edited form as: Cancer Res. 2017 Jun 26;77(16):4460–4471. doi: 10.1158/0008-5472.CAN-17-0015

Figure 5. HuR regulates IDH1 expression in PDAC.

Figure 5

A, RNA sequencing was performed in two PDAC cell lines (HS-766T and MiaPaCa2) modulated by CRISPR/Cas9 against HuR. The heatmap displays transcripts encoding antioxidant enzymes (out of 40 directly involved in ROS detoxification), with significant changes in HuR(−/−) cells as compared with the HuR(+/+) control cells. HuR (ELAVL1, the CRISPR deletion target) is included in the table as reference. B, Schematic of the reversible IDH1 catalytic reaction. NADPH is produced by oxidative decarboxylation. C, HuR and IDH1 mRNA levels in MiaPaca2 CRISPR HuR(+/+) or HuR(−/−) cells; immunoblot analysis of IDH1 and HuR protein in the same cells. D, MiaPaCa2 cells were cotransfected with siRNAs and luciferase reporter constructs (luciferase control or fused with IDH1 3′ UTR). Cells were cultured as indicated for 24 hours. E, αKG levels in MiaPaCa2 cells cultured under the indicated conditions for 24 hours. Error bars, ± SEM of triplicate wells from a representative experiment. *, P < 0.05; **, P < 0.01; ***, P < 0.001. See also Supplementary Figs. S6 and S7.