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. 2019 Mar 1;47(8):3937–3956. doi: 10.1093/nar/gkz128

Figure 2.

Figure 2.

Response of PC-3 cells to pol III inhibitor ML-60218. (A) Mean relative changes in normalized expression of unprocessed pre-tRNATyr and mRNAs encoding CK8, CK14, CK18, SYP, NSE, GRP78, CD55, CD59, CD63 and NANOG in PC-3 cells harvested 48 hrs after treatment with or without 20 μM ML-60218, as determined by RT-qPCR in 3–6 independent experiments. (B) Western blot of NSE, SYP and tubulin in PC-3 cells harvested 48 h after treatment with or without 20 μM ML-60218. (C) Means of PC-3 cell numbers over 3 days with or without 20 μM ML-60218, in six independent experiments. (D) Means of the percentage of PC-3 cells staining positively for Ki67 proliferation marker 2 days after treatment with or without 20 μM ML-60218, in three independent experiments. (E) Western blot of POLR3A, POLR3G, POLR3GL and actin in PC-3 cells harvested 48 h after treatment with or without 20 μM ML-60218. (F) Antiserum to POLR3A was used to immunoprecipitate pol III from PC-3 cells cultured for 48hrs without or with 20 μM ML-60218; panels show western blots of immunoprecipiated protein probed with antibodies to POLR3A, POLR3G and POLR3GL, as indicated. (G) Representative northern blot of U2 snRNA (pol II-transcribed control), unprocessed pre-tRNAiMet and mature tRNAiMet in PC-3 cells harvested 48 hrs after duplicate treatment with or without 20 μM ML-60218. * indicates P < 0.05 relative to control by t-test; ** indicates P < 0.01; *** indicates P < 0.005. Error bars represent S.E.M.