Fig. 5: Glucose starvation specifically facilitates MT-CO2 expression via downregulation of IGF2BP3. | Nature Communications

Fig. 5: Glucose starvation specifically facilitates MT-CO2 expression via downregulation of IGF2BP3.

From: Mitochondrial-cytochrome c oxidase II promotes glutaminolysis to sustain tumor cell survival upon glucose deprivation

Fig. 5

a m6A RIP-qPCR was performed to analyze the levels of m6A-modificated mitochondrial genes in H1299 cells (n = 3 independent experiments). b, c H292 or glucose-starvation-resistant H292 (H292-V) cells were subjected to RNA-seq analyses (b, n = 2 biologically independent samples per experiment) or were subjected to western blot analyses (c). d, e H1299 cells expressing shGFP, shIGF2BP3-1, or IGF2BP3-2 were subjected to western blot (d) or qPCR (e, n  =  3 independent experiments) analyses. f, g H1299 cells expressing IGF2BP3 or vector control (Vec) were subjected to western blot (f) or qPCR (g, n  =  3 independent experiments) analyses. The samples derive from the same experiment but different gels for IGF2BP3, MT-CO2, GAPDH, another for MT-ND1, and another for MT-CO1, MT-ATP6 were processed in parallel (d, f). h, i H1299 cells expressing shGFP, shIGF2BP3-1, or IGF2BP3-2 (h) or H1299 cells expressing IGF2BP3 or Vec (i) were treated with actinomycin D (10 μM) for an indicated time point. Cells were subjected to qPCR analyses. The half-life of the MT-CO2 mRNA levels was shown (n  =  3 independent experiments). j IGF2BP3 RIP-qPCR was performed to analyze the interaction between IGF2BP3 and mitochondrial mRNAs in H1299 cells (n  =  3 independent experiments). k H292-V cells expressing IGF2BP3 or Vec were subjected to western blot analyses. These experiments have been repeated three times with similar results (c, d, f, k). Data were presented as mean ± SD (a, e, g, h–j). Comparisons were performed with two-way ANOVA with Tukey’s (h) or Bonferroni’s (i) test and unpaired two-tailed Student’s t test (a, e, g, j).

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