Figure 4.

JAK2-dependent G6PD Y437 phosphorylation is required for IL-6-induced nucleotide biosynthesis.

(A-G) We stably expressed G6PD shRNA in HSC-3 or CAL-27 cells, and then stably expressed WT Flag-G6PD or Flag-G6PD Y437F. The shRNA targeted the non-coding region of G6PD mRNA, and so that expression of WT Flag-G6PD and Flag-G6PD Y437F is resistant to G6PD shRNA.

(A) Immunoblots were performed using the indicated antibodies. The data are obtained from 3 independent experiments.

(B) Cells were treated with 50 ng/mL IL-6 for 1 h, and immunoprecipitation was performed with an anti-Flag M2 antibody. Immunoblots were performed using the indicated antibodies. The data are obtained from 3 independent experiments.

(C-G) Cells were treated with 50 ng/mL IL-6 for 1 h, and then incubated with [¹⁴C]-glucose (1 μCi, 0.01 mmol/L) for 1 h. The amount of [¹⁴C]-R5P (C), [¹⁴C]-ATP (D), [¹⁴C]-ADP (E), [¹⁴C]-DNA (F) or [¹⁴C]-RNA (G) was measured. The data are presented as the mean ± SD from 3 independent experiments. ∗P < 0.05, ∗∗P < 0.01.