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Fig. 1 | BMC Evolutionary Biology

Fig. 1

From: The evolutionally-conserved function of group B1 Sox family members confers the unique role of Sox2 in mouse ES cells

Fig. 1

Sox2 rescue ability of mouse Sox factors. a Reporter gene assays of mouse Sox factors. The ability to activate or repress the promoter activity via consensus SOX binding motifs was assayed and the relative activities are indicated as ‘fold activation’ on the log scale. A value under 1.0 indicates that the factor functions as a transcriptional repressor. Each assay represents the mean of 3 replicates, error bars mean standard deviations. b Cartoon of Sox2 rescue experiments. 2TS22C ES cells were transfected with an expression vector containing a mouse Sox factor followed by culture without tetracycline (Tc) in FCS-medium to give a pool of transfectants without Sox2. Transfectants were serially passaged in KSR-medium with Tc at a density 3x103/well, and evaluated for their ability to form stem cell colonies. Numbers 1–3 indicate the time points of the evaluation demonstrated in Columns 1–3 of C. c Sox2 rescue ability. The numbers of primary colonies and standard deviation obtained with each Sox gene are indicated under the gene symbol. Column1 shows colonies of primary transfectants grown without Tc. Column 2 shows secondary colonies grown in the presence of Tc. Stem cell colonies were recognized their tightly packed morpohology. Column 3 shows colonies at passage 3 in the presence of Tc if they yielded stem cell colonies at this stage. Stem cell colony formation at this stage indicates rescue of Sox2-null ES cells. d Immunostaining of rescued ES cells. Rescued ES cells at passage 4 in the presence of Tc were stained for Sox2, Oct3/4, Nanog and Klf4. The absence of Sox2 staining with rabbit anti-Sox2 polyclonal antibody in Sox1, Sox3 and Sox15 transfectants confirmed rescue. 2TS22C ES cells cultured with or without Tc for 4 days are shown as positive and negative controls, respectively

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