Figure EV2. The interaction domain between CHK2 and Beclin 1 and characterization of Beclin 1 Ser90 phosphorylation antibody.

• A
  GST‐CHK2 full‐length (FL) and fragments fusion proteins were expressed in bacteria and purified; these proteins were then incubated with in vitro‐translated Flag‐Beclin 1 protein. Western blotting was performed to detect the interaction of Beclin 1 with CHK2.
• B
  GST‐Beclin 1 FL and fragments fusion proteins were expressed in bacteria and purified; these proteins were then incubated with in vitro‐translated Flag‐CHK2 protein. Western blotting was performed to detect the interaction of CHK2 with Beclin 1.
• C
  Cell lysates from 293 cells transfected with WT or mutant forms of Flag‐Beclin 1 were immunoprecipitated with Flag‐M2 beads followed by Western blot using antibody against Beclin 1 Ser90 or Flag.
• D, E
  H1299 cells were pretreated with CHK2 inhibitor II in complete medium for 4 h and then cultured for 1 h in HBSS starvation (D) or H₂O₂ (500 μM) stimulation (E). Western blot detection of p‐CHK2 Thr68, CHK2, p‐Beclin 1 Ser90, p‐Beclin 1 Ser93, and Beclin 1.

Source data are available online for this figure.