Fig. 3: Narta-mediated gene activation is reversible, tunable and specific.
From: Gene activation guided by nascent RNA-bound transcription factors
a Green curves showing the expression level of stdMCP-PH-T2A-GFP controlled by the doxycycline-inducible system at indicated time points. The time to add or remove Dox was indicated by arrows (Day −2 and Day 0, respectively). The blue curves illustrate the protein expression level of exogenous (miniCMV) or endogenous (LMNA and H2B) reporters that were related to the expression of stdMCP-PH-T2A-GFP. The protein expression level was quantified based on fluorescent imaging. Each circle represents the mean intensity of the fluorescent reporter of 100 cells at each time point. Error bar denotes mean ± s.e.m. b–d Representative images (left) and quantifications (right) to show the does-dependent effect of stdMCP-PH on Narta activation of three endogenous reporters. Each dot represents a cell. n = 100 cells. Scale bar, 10 μm. Data are shown as mean ± s.e.m. e Plots to show gene expression levels (log2TPM) in reporter cells (Left: BFPTriTag-LMNA; right: HSPB8-BFPTriTag) transfected with stdPCP-PH (x axis) versus expression in cells transfected with stdMCP-PH (y axis). Mean values of TMP in two replicates were calculated and log2 transformed to show the expression level of each gene. R indicates Pearson’s correlation coefficient, calculated for long-transformed values on all genes except the target gene. Target gene mRNAs are marked in red dots, which are the most significant differentially expressed genes (t-test q value < 0.05 with FDR correction). Source data are provided as a Source Data file.
