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. 2013 Dec 31;8(12):e83426. doi: 10.1371/journal.pone.0083426

Figure 3. Targeting SP1 and/or SP3 via RNAi decreases reportergene activity of the IVS1+1505G reporter gene construct.

Figure 3

miRNA-expressing HIB1b cells were transiently transfected with IVS1+1505G or A, induced and differentiated. During the last 24 hours of differentiation cells were stimulated by a combination of Wy14643 (Wy, 10 µM) and rosiglitazone (rosi, 10 µM). (A) Each cell line expresses two miRNAs targeting either twice SP1, twice SP3, each SP1&SP3 once, UCP1 (Ctrl. U, no transcript detectable in HIB1b cells) or LacZ/shBle (Ctrl. Z, two bacterial genes) The experiment was repeated 8 and 7 times for IVS1+1505G and IVS1+1505A, respectively, each time in triplicates using cells from 2 independent rounds of infection and selection. scram: scrambled shRNA sequence C) Replication of the miRNA experiment using transient transfection of shRNAs with independent sequences. The experiment was carried out 3 times in duplicates. Bars represent mean ± s.d. Stars denote a significant difference from both control vectors for the respective agonist (one way ANOVA for miRNA, Holm-Sidak method, Log transformed data).