Skip to main content
. 2023 Aug 8;13(11):2412–2431. doi: 10.1158/2159-8290.CD-23-0669

Figure 3.

Figure 3. MRTX1719 treatment disrupts PRMT5-regulated processes in MTAP del cancer cells. A, Volcano plots of posttranslational modifications of trypsin-digested peptides from LU99 cells treated for 3 or 5 days with 1 μmol/L MRTX1719 vs. DMSO as measured by LC-MS/MS following antibody enrichment using the SDMA PTMScan kit (top) or the Multi-Pathway PTMScan kit (bottom) that includes phosphorylation (Cell Signaling Technology). B, RNA-seq data were generated from LU99 cells treated with MRTX1719 or DMSO for 3 days, and transcripts were analyzed for alternative RNA splicing using rMATS. Transcripts with retained introns were increased in MRTX1719-treated vs. DMSO-treated cells. C, GSEA was performed on RNA-seq data from 3- and 5-day MRTX1719-treated LU99 cells compared with DMSO-treated. ***, FDR <0.25. D, LU99 cells were treated with 250 nmol/L MRTX1719 for 3 days and analyzed on a Guava flow cytometer. Cell-cycle distribution analysis (left) was performed, and apoptotic cells were analyzed for surface Annexin V–positive, 7-AAD–negative staining (right). Summary data from 2 or 3 independent experiments are presented as the mean ± SD. Statistics were determined using a two-tailed Student t test with significance indicated (*, P < 0.05). E, Cell lysates from LU99 cells treated for 3 days with MRTX1719 and analyzed for selected protein markers by Western blot.

MRTX1719 treatment disrupts PRMT5-regulated processes in MTAP del cancer cells. A, Volcano plots of posttranslational modifications of trypsin-digested peptides from LU99 cells treated for 3 or 5 days with 1 μmol/L MRTX1719 vs. DMSO as measured by LC-MS/MS following antibody enrichment using the SDMA PTMScan kit (top) or the Multi-Pathway PTMScan kit (bottom) that includes phosphorylation (Cell Signaling Technology). B, RNA-seq data were generated from LU99 cells treated with MRTX1719 or DMSO for 3 days, and transcripts were analyzed for alternative RNA splicing using rMATS. Transcripts with retained introns were increased in MRTX1719-treated vs. DMSO-treated cells. C, GSEA was performed on RNA-seq data from 3- and 5-day MRTX1719-treated LU99 cells compared with DMSO-treated. ***, FDR <0.25. D, LU99 cells were treated with 250 nmol/L MRTX1719 for 3 days and analyzed on a Guava flow cytometer. Cell-cycle distribution analysis (left) was performed, and apoptotic cells were analyzed for surface Annexin V–positive, 7-AAD–negative staining (right). Summary data from 2 or 3 independent experiments are presented as the mean ± SD. Statistics were determined using a two-tailed Student t test with significance indicated (*, P < 0.05). E, Cell lysates from LU99 cells treated for 3 days with MRTX1719 and analyzed for selected protein markers by Western blot.