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. 2019 Sep 18;11(9):1394. doi: 10.3390/cancers11091394

Figure 3.

Figure 3

Interaction between Janus kinase 2 (JAK2) and Forkhead box O3 (FOXO3) protein in A498 and ACHN. (A) Silencing IL4Rα in A498 and ACHN cells reduced the interaction between JAK2 and FOXO3. Cells were transfected with control siRNA or siRNA against IL4Rα, and then cell lysates were immunoprecipitated with the anti-pJAK2 antibody. The immunoprecipitated proteins were resolved on the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotted by anti-IL4Rα, IL13Rα1, FOXO3, and pJAK2 antibody. The light chain of IgG was used for the loading control. (B) 293T cells were co-transfected with HA- JAK2 and Flag-FOXO3 (O.E.) or a control plasmid DNA (pCMV3-C-HA and pECE, Con.) as indicated. Then cell lysates were immunoprecipitated with anti-Flag, FOXO3, HA, JAK2, or pTyr antibody. The immunoprecipitated proteins were resolved on the SDS-PAGE and immunoblotted by the indicated antibody, respectively. The light chain of IgG and Coomassie Blue staining of SDS-PAGE were used for the loading control. (C) Silencing JAK2 in A498 and ACHN cells increased the expression of FOXO3. Cells were transfected with control siRNA or siRNA against JAK2, and then cell lysates were resolved on the SDS-PAGE and immunoblotted by anti-FOXO3 and JAK2 antibody. β-actin was used for the loading control. (D) Pharmacological inhibition of JAK2 in A498 and ACHN cells increased the expression of FOXO3. Cells were treated with dimethyl sulfoxide (DMSO) vehicle control or the indicated concentration of AZD1480, and then cell lysates were resolved on the SDS-PAGE and immunoblotted by anti-FOXO3, pJAK2, and JAK2 antibody. β-actin was used for the loading control.