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. Author manuscript; available in PMC: 2012 Nov 1.
Published in final edited form as: J Cell Physiol. 2011 Nov;226(11):3004–3013. doi: 10.1002/jcp.22648

Figure 1. TGFβ and bleomycin activates Akt-mTOR pathway and synthesis of ECM proteins in a dose- and time-dependent manner.

Figure 1

(A) NIH3T3 fibroblasts were grown to confluence, treated with TGFβ (0, 25, 50, 100 and 250pM) in serum free medium and further incubated at 37°C in a CO2 incubator. After 6, 12 and 24 hours, conditioned media was collected and cells were lysed using RIPA buffer with protease and phosphatase inhibitors. (B) NIH3T3 fibroblasts were grown to confluence, treated with Bleomycin (0, 1, 2.5 and 5mU) in serum free medium and further incubated at 37°C in a CO2 incubator. After 6, 12 and 24 hours, conditioned media was collected and cells were lysed using RIPA buffer. Both the conditioned media and cell lysates were subjected for western analysis for fibronectin and the cell lysates were further analyzed for changes in phosphorylation of signaling molecules in the Akt-mTOR pathway.