Fig. 3.
CARM1 and GRIP1 activate MMP-9 promoter transcription and CARM1 methyltransferase activity is required. (A) Schematic diagram of CARM1 and CARM1E267Q. E: glutamic acid; Q: glutamine. (B) HeLa cells were transiently transfected with the MMP-9-Luc reporter construct and 0 to 0.6 µg of pSG5-CARM1 or pSG5-CARM1E267Q, and MMP-9 promoter activity determined as described in Materials and Methods. The results are the mean ± S.E. of at least three independent experiments. * p ≤0.05 compared to pSG5 only transfected PMA treated samples; # p ≤0.05 between the two linked samples; NS, not significant compared to pSG5 only transfected PMA treated samples. (C) HeLa cells were transiently transfected with 0.6 µg of pSG5.HA, pSG5.HA-CARM1 or pSG5.HA-CARM1E267Q for 48 h and whole cell lysates isolated and subjected to immunoblotting analysis. Actin was utilized as a loading control. (D) HeLa cells were transiently transfected with the MMP-9-Luc reporter construct and 0 to 0.6 µg of pSG5-GRIP1, and MMP-9 promoter activity determined as described in Materials and Methods. The results are the mean ± S.E. of at least three independent experiments. * p ≤0.05 compared to pSG5 only transfected PMA treated samples; NS, not significant compared to pSG5 only transfected PMA treated samples. (E) HeLa cells were transiently transfected with 0.6 µg of pSG5.HA or pSG5.HA-GRIP1 for 48 h and whole cell lysates subjected to immunoblotting analysis. Actin was utilized as a loading control.
