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. 2017 Nov 13;127(12):4498–4515. doi: 10.1172/JCI91553

Figure 10. Lnc-BM modulates kinase activity of JAK2.

Figure 10

(A) Kinase assay using anti-JAK2 immunoprecipitates retrieved from 231-Br cells transfected with indicated siRNAs followed by OSM treatment. (B and C) In vitro kinase assay using recombinant JAK2 JH1/JH2 domain (B) or JAK2 JH1 domain (C), GST-STAT3, and indicated RNA transcripts in the presence and absence of ATP. (D) Graphic illustration of Lnc-BM–mediated JAK2 JH1/JH2 domain conformational alteration from autoinhibition to activation. (E) FLAG-tag pull-down followed by IB detection using antibodies in HEK-293T cells transfected with indicated vectors. (F) Anti-JAK2 immunoprecipitates were detected using indicated antibodies in 231-Br cells transfected with indicated siRNA. (G) Kinase assay using anti-FLAG immunoprecipitates in γ-2A cells transfected with SFB-JAK2 (aa 482–809), in the presence of indicated RNA transcripts and ATP.