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. 2017 May 2;45(13):7681–7696. doi: 10.1093/nar/gkx364

Figure 4.

Figure 4.

Comparison of transactivation by c-Myb in WT and D152V mutant form. (A) CV-1 cells were transfected with c-Myb responsive reporter plasmids (pGL4b-3xMRE(GG)-myc or pmim3mim) and plasmids encoding full-length HA-tagged c-Myb or c-Myb D152V. The reporter activations are presented as relative luciferase units. The western blots were analysed using anti-HA (c-Myb) and anti-GAPDH primary antibodies. (B) CV-1 cells were transfected with c-Myb responsive reporter plasmids (pGL3-MYC, pGL3-STAT5A or pGL3-LMO2) and plasmids encoding full-length HA-tagged c-Myb or c-Myb D152V. The reporter activations are presented as relative luciferase units. (C) Upper panel: HD-11 cells were transfected with plasmids encoding full-length HA-tagged c-Myb or c-Myb D152V. Total RNA was isolated and mim-1 expression was measured by qRT-PCR. Lower panel: HD-11 cells were transfected with the c-Myb responsive reporter plasmid pmim3mim and the same c-Myb plasmids as above. The reporter activations are presented as relative luciferase units. The western blots were analysed using anti-HA (c-Myb) and anti-GAPDH primary antibodies. All luciferase and qRT-PCR results are presented as mean ± SD of three independent biological replicates, each performed in triplicates (n = 3). Significance was evaluated by unpaired, two-tailed t-tests on selected pairs.