American Society of Hematology

Figure 2.

$Figure 2. In-depth quantitative proteomics of MCL cell lines treated with HSP90 inhibition. (A) Experimental strategy. Lysates from MCL cell lines Mino, JeKo-1, and MAVER-1 (parental and/or transduced to express either wt BTK or BTK C481S) treated with either DMSO or AUY922 50 nM for 24 hours were used for protein isolation and digestion. Peptides were labeled with Tandem Mass Tag 10-plex (TMT10plex) reagents and peptide fractionation. Multiplexed quantitative MS data were collected and analyzed. (B) Comparison between protein levels in parental and transduced Mino, JeKo-1, and Maver-1 cell lines for DMSO- vs AUY922-treated cells. Proteins downregulated and upregulated by log2 fold ≥0.5 are represented in red. (C) Volcano plot demonstrating the distribution of protein fold changes between parental and transduced Mino, JeKo-1, and Maver-1 cell lines for AUY922- vs DMSO-treated cells, highlighting the most highly upregulated and downregulated proteins following HSP90 inhibition with AUY922. (D) Pathway analysis using Ingenuity pathway analysis (Qiagen) identifying the cellular and molecular pathways most highly affected by protein level changes following HSP90 inhibition with AUY922 in MCL lines. LC, liquid chromatography.$

In-depth quantitative proteomics of MCL cell lines treated with HSP90 inhibition. (A) Experimental strategy. Lysates from MCL cell lines Mino, JeKo-1, and MAVER-1 (parental and/or transduced to express either wt BTK or BTK C481S) treated with either DMSO or AUY922 50 nM for 24 hours were used for protein isolation and digestion. Peptides were labeled with Tandem Mass Tag 10-plex (TMT10plex) reagents and peptide fractionation. Multiplexed quantitative MS data were collected and analyzed. (B) Comparison between protein levels in parental and transduced Mino, JeKo-1, and Maver-1 cell lines for DMSO- vs AUY922-treated cells. Proteins downregulated and upregulated by log2 fold ≥0.5 are represented in red. (C) Volcano plot demonstrating the distribution of protein fold changes between parental and transduced Mino, JeKo-1, and Maver-1 cell lines for AUY922- vs DMSO-treated cells, highlighting the most highly upregulated and downregulated proteins following HSP90 inhibition with AUY922. (D) Pathway analysis using Ingenuity pathway analysis (Qiagen) identifying the cellular and molecular pathways most highly affected by protein level changes following HSP90 inhibition with AUY922 in MCL lines. LC, liquid chromatography.

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