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. 2017 Jun 23;6:e22477. doi: 10.7554/eLife.22477

Figure 5. MPI loss suppresses the Warburg effect in embryonic cells.

(A) Heat map based on RNA-seq data analysis showing downregulation of glycolytic genes in mpi morphants at 24 hpf. Standardized fold change (FC) in mpi morphants in WT and p53 MT was calculated relative to standard control morpholino (std MO)-injected embryos. The color scheme represents gene expression changes in a log2 scale, in the range of −1.5 (blue, decreased) to 1.5 (red, increased). (B) qPCR analysis showed decrease of Pfk and Ldha mRNA levels in siMpi MEFs compared to NC. The Ct values were normalized relative to the expression of Rps28 gene. Relative fold change was calculated as delta Ct in siMpi compared to NC. (C) 2-Deoxyglucose uptake measurement demonstrated a decrease in siMpi MEFs compared to the NC. Normalized relative % change is shown. (D) Measurement of lactate levels in MEFs following siMpi-mediated depletion of Mpi was assessed by lactate assay and showed a significant reduction of lactate in siMpi cells. Results were normalized to MTT assay. Relative amounts of lactate are shown (% of NC). (E) Measurement of lactate levels in 4 dpf zebrafish embryos from mpimss7/+ MT incrosses showed a decrease in lactate production compared to the offspring of the mpi+/+ incrosses. Results were normalized to number of embryos. (F) Measurement of lactate levels in 4 dpf zebrafish embryos injected with std MO or mpi MO showed significant decrease in lactate production in mpi MO-injected embryos compared to std MO injected WT embryos, but no decrease when injected in to p53 MT embryos. Results were normalized to number of embryos. Two-tailed Student's t-test was applied in panels B-F.

DOI: http://dx.doi.org/10.7554/eLife.22477.014

Figure 5—source data 1. Changes in expression of the genes involved in glycolysis.
Partial dataset of RNA-seq analysis performed in 24 hpf zebrafish embryos injected with std or mpi MO. Log2 fold change (FC) values relative to WT + std MO. The list of showing the changes in expression of the genes involved in glycolysis.
DOI: http://dx.doi.org/10.7554/eLife.22477.015
elife-22477-fig5-data1.xlsx (38.9KB, xlsx)
DOI: 10.7554/eLife.22477.015

Figure 5.

Figure 5—figure supplement 1. Loss of MPI leads to decrease cell viability in embryonic and cancer cells.

Figure 5—figure supplement 1.

Cell quantification in MEFs (A) and SJSA cells (B) with and without siMPI transfection. (C) Cell proliferation in siMpi MEFs was assessed by MTT assay. Relative amounts of mitochondrial activity are shown (% of NC). p-Value using two-tailed paired Student’s t-test.
DOI: http://dx.doi.org/10.7554/eLife.22477.016
Figure 5—figure supplement 2. Loss of MPI leads to inhibition of glycolysis in HCT116 cancer cells.

Figure 5—figure supplement 2.

(A) MPI activity after MPI knockdown with siMPI in HCT116 WT and p53 null colon cancer cells compared to those transfected with control siRNA (NC). (B) qPCR demonstrated downregulation of glycolytic genes in HCT116 cells transfected with siMPI compared to NC. The Ct values were normalized to expression of Rps18 gene. Relative fold change was calculated as delta Ct in siMPI compared to NC. (C) Lactate levels measured in WT and p53 null HCT116 cells transfected with siMPI compared to NC. (D) ATP levels measured in HCT116 cells transfected with siMPI or NC showed decreased ATP following MPI silencing in siMPI compared to NC. All p-values calculated using paired two-tailed Student’s t-test.
DOI: http://dx.doi.org/10.7554/eLife.22477.017