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. Author manuscript; available in PMC: 2013 Aug 15.
Published in final edited form as: Free Radic Biol Med. 2012 Jun 23;53(4):974–982. doi: 10.1016/j.freeradbiomed.2012.06.019

Figure 2. miR-302 expression is repressed in quiescence compared to proliferating cells.

Figure 2

Normal human fibroblasts were induced to quiescence by contact inhibition. Exponential cultures were collected 24 h after replating of contact inhibited quiescent cultures. (A) RNA samples from quiescent and proliferating fibroblasts were isolated and analyzed for miR-302 abundance as well as mRNA levels of ARID4a and CCL5. Fold change was calculated relative to RNA levels in exponential cultures. Asterisks represent statistical significance relative to exponential cultures. (B) Protein samples from quiescent and proliferating cells were isolated and immunoblotting for ARID4a was performed. Actin levels were used for comparison. (C) An ELISA assay was used to measure CCL5 in media collected from exponential and quiescent fibroblasts. Asterisks represent statistical significance relative to exponential cultures. (D) RNA samples from low and high density monolayer cultures of FaDu cells were assayed for miR-302, ARID4a, and CCL5. Asterisks represent statistical significance relative to low density cultures. (E) An inverse correlation between miR-302 and ARID4a was obtained when the delta Ct of ARID4a (closed circles) and miR-302 (open circles) were plotted relative to the percentage of S-phase. n = 3, error bars represent standard deviation.