Skip to main content
. 2011 Jul;31(14):2854–2866. doi: 10.1128/MCB.05397-11

Fig. 1.

Fig. 1.

CIITA interacts with myogenin. (A) Isolation of myogenin-interacting proteins by a GST affinity approach. Nuclear differentiated C2C12 extract (45 mg) was incubated with the indicated constructs. Following incubation, the glutathione beads were washed and interacting proteins eluted with increasing amounts of salt. SDS-PAGE gels were silver stained. The mobility of the GST fusion proteins is marked with a black asterisk. The box indicates the region that was excised and trypsin digested for protein identification by liquid chromatography/mass spectrometry (LC/MS). (B) CIITA interacts with myogenin. HEK cells were transiently transfected with myc-CIITA and EMSV-myogenin and cell extracts immunoprecipitated with antibodies against CIITA or myogenin. (C) CIITA does not interact with MyoD. HEK cells were transiently transfected with myc-CIITA and EMSV-MyoD and cell extracts immunoprecipitated with antibodies against CIITA or MyoD. (D) CIITA does not interact with Myf5. HEK cells were transiently transfected with myc-CIITA and EMSV-Myf5 and cell extracts immunoprecipitated with antibodies against CIITA or Myf5. (E) HEK cells were transiently transfected with myc-CIITA and EMSV-Myf6, and cell extracts were immunoprecipitated with antibodies against CIITA or Myf6. (F) Endogenous interaction of CIITA with myogenin in myotubes. Extracts from C2C12 cells differentiated for 2 days were immunoprecipitated with antibodies against myogenin (F5D), and the blot was probed with antibodies against CIITA (7-1H). The blot was then stripped and reprobed with antibodies against myogenin. For each panel, the antibodies used for the immunoprecipitation and the probe are labeled above the lanes. The immunoprecipitated sample is labeled B (bound fraction) and the supernatant, or unbound fraction, is labeled UB. The lysate used for the immunoprecipitation is labeled EXT (extract). Reciprocal blots are shown. The blots were also probed with the antibody used for the immunoprecipitation to confirm that the target protein was immunoprecipitated in each case (data not shown).