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. 2020 Aug 19;9:e59166. doi: 10.7554/eLife.59166

Figure 5. PKM2 is not required for transcription of HIF1α and STAT5α target genes in NK cells.

Figure 5.

(a–c) Pkm2NK-WT and Pkm2NK-KO cultured and purified cells were stimulated with (IL-2/12) or left unstimulated (low-dose IL-15) for 18 hr. HiSeq RNA sequencing was then performed. (a) Differential gene expression analysis was carried out to assess total gene changes between IL-2/12 stimulated Pkm2NK-WT and Pkm2NK-KO cells. Total gene changes were assessed at a fold change cut-off of 2 and a p value of 0.05 with an FDR of 0.05. (c) Expression levels of key HIF1α target genes were assessed and compared between Pkm2NK-WT and Pkm2NK-KO cells (c) Expression levels of key STAT5 target genes were assessed and compared between Pkm2NK-WT and Pkm2NK-KO cells. (d-f) Cultured wildtype NK cells were stimulated with IL-2/12 +/- TEPP-46/vehicle for 18 hr. HiSeq RNA sequencing was then performed. (d) Differential gene expression analysis was carried out to assess total gene changes between IL-2/12 TEPP-46 (50 μM) and IL-2/12 Vehicle (0.1% v/v DMSO). Total gene changes were assessed at a fold change cut-off of 2 and a p value of 0.05 with FDR 0.05. (e) Expression levels of key HIF1α target genes were assessed and compared between IL-2/12 TEPP-46 (50 μM) and IL-2/12 Vehicle (0.1% v/v DMSO) treated cells. (f) Expression levels of key STAT5 target genes were assessed and compared between IL-2/12 TEPP-46 (50 μM) and IL-2/12 Vehicle (0.1% v/v DMSO) treated cells. Data are from n = 3 biological replicates per group and are displayed as mean values. (b–c) RPKM values were normalised to the average of PKM2NK-WT unstimulated. Fold change values were then log transformed and displayed in heat maps. (e–f) RPKM values were normalised to the mean of both groups combined and then converted to fold change from the mean. Data were then log transformed and displayed as heat maps. n = 3 mice per group. Data are mean and were analysed by (a–c) two-way ANOVA with multiple comparisons or (d-f) one-way ANOVA with Tukey post-test. ns – not significant *p>0.05.