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. Author manuscript; available in PMC: 2023 May 24.
Published in final edited form as: Oncogene. 2023 Apr 7;42(21):1716–1727. doi: 10.1038/s41388-023-02688-5

Figure 4. SREBP2 and c-Myc are required for RON-mediated upregulated of CBS and glycolysis, respectively.

Figure 4.

(A) Enriched transcription factor signatures based on differentially expressed genes from RON-modulated T47D cells (Enrichr TRANSFAC); c-MYC/MAX and SREBF2 (SREBP2) are emphasized by red boxes. (B) Relative SREBP2 gene expression determined by qRT-PCR analysis on samples from RON expressing R7 cells (R7 Control; n=3 biological replicates) versus isogenic RON-deplete counterparts (R7 sgRON; n=3 biological replicates) with and without RON inhibition through 5 μM BMS777607 treatment for 6 hours. (C) Western blot analysis of R7 sgRON cells with or without SREBP2 overexpression (SREBP2 OE) examining cleaved SREBP2 levels and CBS target gene levels (LSS, FDFT1), c-Myc, and glycolysis gene levels (HK2, LDHA) with actin used as a loading control. (D) Mammosphere formation of R7 sgRON cells with or without SREBP2 OE (n=3 biological replicates). (E) Western blot analysis of c-Myc expression in RON-modulated MCF7, T47D, and R7 cells with actin as a loading control. (F) Western blot analysis of R7 cells with or without c-Myc knockdown by shRNA (shMyc) and R7 sgRON cells with c-Myc overexpression (c-Myc OE) examining c-Myc levels, glycolysis (HK2, LDHA), cleaved SREBP2, and CBS genes (FDFT1, LSS) with actin as a loading control. (G) Mammosphere formation of R7 cells with or without shMyc (n=3 biological replicates). (H) Mammosphere formation of R7 sgRON cells with or without c-Myc OE (n=3 biological replicates). Western blot expression comparisons are only intended to be made within individual boxes. Data displayed represent mean values ± SD. ns, not significant; *P<0.05; **P<0.01; ***P<0.001.