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. 2013 Sep 2;8(9):e73210. doi: 10.1371/journal.pone.0073210

Figure 6. Effect of c-fos on aggrecan (Agg) and collagen type II (Col2) expression and cell proliferation in NP cells.

Figure 6

(A) NP cells or stable AP-1 reporter cell were cotransfected with the Agg (400 ng) or Col2 (400 ng) reporter constructs and increasing concentrations of the WT-c-fos plasmid (100–500 ng), the DN-c-fos (100–500 ng) plasmid, or empty backbone. *P < 0.05 indicates differences between groups. Error bars represent the SD. (B) Detection of Agg protein expression by immunofluorescence microscopy. NP cells were cultured with (a) or without (b) WT-c-fos for 24 h, fixed, and stained with an antibody against Agg (green). Scale bar= 100 μm(C) Real-time-RT-PCR analysis of c-fos, Agg or Col2 expression in cells treated with vehicle (control) or a WT-c-fos expression plasmid. Total RNA was extracted from transfected cells and real-time RT-PCR was used to determine c-fos, Agg, Col2 and GAPDH RNA levels as an internal control. (D) The NP cells were treated with c-fos (1 ng/mL) in 96-well plates. Cell proliferation was evaluated using the MTT viability assay 12 or 24 h after treatment. n.s. = not significant.