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. 2015 Jan 20;116(3):408–417. doi: 10.1002/jcb.24989

Figure 1.

Figure 1

Differential binding of nuclear proteins to COL1A2 promoter probes. (A) Quantitative Light Cycler RT‐PCR showing downregulation of the human COL1A2 gene in SV‐WI38 compared to wildtype WI38 cells which was set as 100%. Experiments were performed in triplicates and normalised to β‐actin and HSPC3 expression, P < 0.001. (B) Western Blot confirming decreased type I collagen protein in SV‐WI38 cells; GAPDH served as loading control. (C) Probes used in EMSAs. The probe (−100/−61) represents the COL1A2 promoter region from −100 to −61 relative to the transcriptional start site that contains an inverted CCAAT Binding Element (CBE) and an adjacent Collagen Modulating Element (CME). The probe (3xCME) consists of the CME consensus sequence repeated three times. (D) EMSA showing the binding of SV‐WI38 nuclear proteins to either the (−100/−61) probe or the (3xCME) probe used either single‐stranded (s = sense or as = antisense), or double‐stranded (DS), respectively. Note that the position of the free probe differs between single‐stranded and double‐stranded oligonucleotides.