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. 2019 Feb 15;11(2):964–973.

Figure 5.

Figure 5

Phosphorylation of GSK3β at serine 9 is required for the inhibitory effect of BNP on TIMP2 expression and the ensuing effect on ECM accumulation. Immortalized murine podocytes were subjected to transient transfection with plasmid vectors encoding the empty vector (EV) or a hemagglutinin (HA)-conjugated nonphosphorylatable GSK3β mutant, in which the serine 9 residue is replaced by alanine (S9A). Cells were treated with TGFβ1 (10 ng/ml) in the presence or absence of BNP (10-6 M) for 48 h. A. Cells lysates were processed for immunoblot analysis for TIMP2, HA, GSK3β and actin (full-length blots are included in the Figure S3). B. Cell cultures were stained with Sirius red and visualized under light microscope. Representative microscopic images were shown. Bar =100 µm. C. Cell lysates were collected and processed for Sirius red total collagen assay. Relative abundance of total collagen is shown. *P < 0.05 versus TGFβ1 alone-treated Control cells (n=3).