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Figure 4 | Journal of Neuroinflammation

Figure 4

From: Calmodulin kinase II-dependent transactivation of PDGF receptors mediates astrocytic MMP-9 expression and cell motility induced by lipoteichoic acid

Figure 4

Involvement CaMKII in LTA-induced MMP-9 expression in RBA-1. (A, B) Cells were pretreated with inhibitors of calmodulin (CaMI) or CaM kinase II (KN-62) for 1 h and then incubated with 50 μg/ml LTA for 24 h. Conditioned media were collected and analyzed by gelatin zymography. Cell lysates were assayed by western blot using an anti-GAPDH antibody as a control. (C) Time dependence of LTA-stimulated phosphorylation of CaMK II. RBA-1 cells were incubated with 50 μg/ml LTA for the indicated time intervals. (D) Cells were pretreated CaMI or KN-62 for 1 h and then incubated with LTA for the indicated time intervals. Cell lysates were assayed by western blot using an anti-phospho-CaMKII antibody. The membranes were stripped and re-probed with anti-GAPDH antibody as a control. (E) Cells were transfected with a CaMKII shRNA plasmid for 48 h, and then incubated with LTA for 24 h. Cell lysates were assayed by western blot using an anti-CaMKII or anti-GAPDH (as a control) antibody. Conditioned media were collected and analyzed by gelatin zymography. Data are expressed as mean ± SEM (A-C) or mean (D) of three independent experiments (n = 3). *P < 0.05; #P < 0.01, as compared with the cells exposed to LTA alone. The figure represents one of at least three individual experiments.

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