Skip to main content
Fig. 7 | Journal of Experimental & Clinical Cancer Research

Fig. 7

From: A novel protein encoded by ZCRB1-induced circHEATR5B suppresses aerobic glycolysis of GBM through phosphorylation of JMJD5

Fig. 7

The effects of JMJD5 phosphorylated by HEATR5B-881aa on glycolysis and proliferation in GBM. A The colocalization of HEATR5B-881aa and JMJD5 in U251 and U373 cells by IF assays. Green, HEATR5B-881aa; red, JMJD5; blue, DAPI nuclear staining. Scale bars, 10 μm. B Lysates of U251 cells were subjected to immunoprecipitation (IP) and immunoblotting (IB) with HEATR5B-881aa and JMJD5 antibodies. C Lysates of 293 T cells transfected with FLAG-HEATR5B-881aa and GST-JMJD5 plasmids were subjected to IP and IB with FLAG tag and GST tag antibodies. D The direct interaction between HEATR5B-881aa and JMJD5 was confirmed by GST pull-down assays. GST protein functioned as a negative control. E Left panel, in vitro kinase assays were performed and detected by autoradiography (arrow, phosphorylated band). Right panel, proteins were visualized by Coomassie brilliant blue (CBB) staining. F The phosphorylated bands were subjected to mass spectrometry and S361 was identified. G In vitro kinase assays and CBB staining were conducted after S361 mutation. H The effects of S361 phosphorylation on JMJD5 stability were detected by Cycloheximide (CHX) chase assays. Data are presented as the mean ± SD (n = 3, each group). *P < 0.05, **P < 0.01 vs. JMJD5-WT group by one-way ANOVA. I-K The effects of JMJD5 phosphorylation on glucose consumption (I), lactate production (J), and proliferation (K) in U251 and U373 cells. Data are presented as the mean ± SD (n = 3, each group). *P < 0.05, **P < 0.01, ***P < 0.001 vs. JMJD5-WT group by one-way ANOVA. L The effects of JMJD5 phosphorylation on ECAR in U251 and U373 cells. M Subcutaneous xenograft nude mouse models using U251 and U373 cells transfected with JMJD5-WT, JMJD5-S361A, or JMJD5-S361E were established with tumor growth curves. Data are presented as the mean ± SD (n = 5, each group). **P < 0.01, ***P < 0.001 vs. JMJD5-WT group by two-way ANOVA

Back to article page