Figure 3.

TLR agonists induce upregulation of PPM1A expression. (A) THP-PPM1A^prom cells were treated with PMA (100 ng/ml) to induce macrophage differentiation. At the indicated time points post differentiation, cells were analyzed by flow cytometry to measure the amount of GFP production as a function of PPM1A promoter activity. MFI: mean fluorescence intensity of GFP signal. Results represent the means ± S.D. of three independent experiments. *p < 0.01 relative to day 0 control cells. (B) PMA-differentiated THP-PPM1A^prom cells were left untreated, infected with Mtb, or stimulated with imiquimod (5 µg/ml) or Pam3CSK4 (300 ng/ml) for 48 h. Flow cytometric analysis was used to measure the amount of GFP production as a function of PPM1A promoter activity. MFI: mean fluorescence intensity of GFP signal. (C) Cell lysates from GM-CSF-differentiated hMDMs treated with imiquimod (5 µg/ml) or Pam3CSK4 (300 ng/ml) for 48 h were prepared and used to analyze PPM1A protein levels by Western blotting. The untreated lane was cropped from the same gel/blot for easier comparison to the experimental lanes as the original blot contained irrelevant lanes in between the untreated and treated samples. The full-length/uncropped blot is presented in Supplementary Fig. 3.