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. 2020 Jun 24;136(22):2557–2573. doi: 10.1182/blood.2020005795

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Figure 2. — Knocking out MIF exacerbates Cfz-induced mitochondrial dysfunction in MM cells. (A) The top toxicologic and dysregulated cellular pathways predicted by Ingenuity Pathway Analysis based on differentially expressed genes in CTR-KO or MIF-KO ARP-1 or MM.1S MM cells. The circle surface area is proportional to ratio, and the color intensity of circles indicates -log (P value). Flow cytometry histogram (B) and summarized results (C) of the tetramethylrhodamine ethyl ester (TMRE) fluorescence of CTR-KO or MIF-KO ARP-1 or MM.1S MM cells after pulse treatment with DMSO or Cfz for 14 hours. (D) Pie chart showing the percentage of fatty oxidation disorder–related metabolites (blue) among the top 22 upregulated metabolites in Cfz-treated MIF-KO ARP-1 MM cells. (E) Heatmap showing the fold change (FC) of fatty oxidation disorder–related metabolites in Cfz-treated MIF-KO vs CTR-KO ARP-1 MM cells. Oxygen consumption rates (OCRs) for mitochondrial respiration function in CTR-KO or MIF-KO MM.1S (F), ARP-1 (G), KMS-11 or KMS-11/Cfz (L) MM cells were measured by using the Seahorse XF Cell Mito Stress Test, which consists of automated treatment with oligomycin, carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP), and the combination of antimycin A and rotenone at the indicated times using the XF Extracellular Flux Analyzer. Summarized results of basal OCR, maximum OCR, spare OCR (H,M), or ATP production OCR (I,N) are shown for 3 independent replicates as described in panels F, G, and L. Summarized results showing basal OCR, maximum OCR, and spare OCR in DMSO- or Cfz-treated MM.1S (J) or ARP-1 MM (K) cells. For panel B, one representative result of at least 3 independent experiments is shown. The Student t test was used to compare 2 samples. *P < .05; **P < .01. n.s., not significant.

Figure 2. — Knocking out MIF exacerbates Cfz-induced mitochondrial dysfunction in MM cells. (A) The top toxicologic and dysregulated cellular pathways predicted by Ingenuity Pathway Analysis based on differentially expressed genes in CTR-KO or MIF-KO ARP-1 or MM.1S MM cells. The circle surface area is proportional to ratio, and the color intensity of circles indicates -log (P value). Flow cytometry histogram (B) and summarized results (C) of the tetramethylrhodamine ethyl ester (TMRE) fluorescence of CTR-KO or MIF-KO ARP-1 or MM.1S MM cells after pulse treatment with DMSO or Cfz for 14 hours. (D) Pie chart showing the percentage of fatty oxidation disorder–related metabolites (blue) among the top 22 upregulated metabolites in Cfz-treated MIF-KO ARP-1 MM cells. (E) Heatmap showing the fold change (FC) of fatty oxidation disorder–related metabolites in Cfz-treated MIF-KO vs CTR-KO ARP-1 MM cells. Oxygen consumption rates (OCRs) for mitochondrial respiration function in CTR-KO or MIF-KO MM.1S (F), ARP-1 (G), KMS-11 or KMS-11/Cfz (L) MM cells were measured by using the Seahorse XF Cell Mito Stress Test, which consists of automated treatment with oligomycin, carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP), and the combination of antimycin A and rotenone at the indicated times using the XF Extracellular Flux Analyzer. Summarized results of basal OCR, maximum OCR, spare OCR (H,M), or ATP production OCR (I,N) are shown for 3 independent replicates as described in panels F, G, and L. Summarized results showing basal OCR, maximum OCR, and spare OCR in DMSO- or Cfz-treated MM.1S (J) or ARP-1 MM (K) cells. For panel B, one representative result of at least 3 independent experiments is shown. The Student t test was used to compare 2 samples. *P < .05; **P < .01. n.s., not significant.