Protein and mRNA expression assay of Ultramarine and ShadowR fusion constructs. (a) HeLa cells expressing tandem constructs of mEGFP, mRuby2, and mScarlet with Ultramarine or ShadowR were lysed and immunoblotted with anti-FLAG and anti-β-actin antibodies. (b–d) Quantitative analysis of immunoblotting. The band intensities of FLAG-tagged fluorescent and chromo proteins were divided by those of actin. Error bars indicate S.E.M. for seven independent experiments. Asterisks denote statistical significance (t test, *P < 0.05, **P < 0.01, ***P < 0.001, N.S. = not significant). (e) The expression of indicated genes in HeLa cells was quantified with qRT-PCR. As an endogenous reference gene, beta-actin mRNA levels were measured. Asterisks denote statistical significance (t test, *P < 0.05, **P < 0.01, ***P < 0.001, N.S. = not significant).