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. 2018 Aug 3;103(12):2049–2058. doi: 10.3324/haematol.2018.191684

Figure 2.

Figure 2.

OTX015 modulates microRNA-96-5p expression in diffuse large B-cell lymphoma models. (A) OTX015 upregulates miR-96-5p in a time-dependent manner. Two germinal center B-cell (GCB)-diffuse large B-cell lymphoma (DLBCL) cell lines (DOHH-2, OCI-LY-1) and two activated B-cell (ABC)-DLBCL cell lines (SU-DHL-2, HBL-1) were treated with dimethyl sulfoxide (DMSO) or 500 nM OTX015 for 4, 24, and 48 h. Expression of miR-96-5p was determined by TaqMan quantitative reverse transcription polymerase chain reaction (qRT-PCR). Expression of RNU6B was used for normalization. For each time-point, the mean fold-change relative to the DMSO control is shown. (B) OTX015 treatment of DLBCL cells downregulates PRMT5. Two GCB-DLBCL (DOHH-2, OCI-LY-1) and two ABC-DLBCL (SU-DHL-2, HBL-1) cell lines were treated with DMSO or 500 nM OTX015 for 4, 24, and 48 h. Expression of PRMT5 was determined by qRT-PCR. GAPDH expression was used for normalization. For each time-point, the mean fold-change relative to the DMSO control is shown. (C) OTX015 reduces PRMT5 protein levels in DOHH-2 and SU-DHL-2 cells treated with DMSO or 500 nM OTX015. GAPDH was used as a loading control. PRMT5 signals were quantified using ImageJ (http://rsb-web.nih.gov/ij/) and normalized to GAPDH signals. Representative images of two independent Western blot analyses are shown. The graphs show the mean of three independent experiments. **P<0.01. Error bars denote the standard error.