Fig. 7: Recovery of reprogramming capacity in immortalized DIDO3ΔE16 MEF, and model for transcription termination in WT and DIDO3ΔE16 cells. | Cell Death & Disease

Fig. 7: Recovery of reprogramming capacity in immortalized DIDO3ΔE16 MEF, and model for transcription termination in WT and DIDO3ΔE16 cells.

From: Impaired stem cell differentiation and somatic cell reprogramming in DIDO3 mutants with altered RNA processing and increased R-loop levels

Fig. 7

a Scheme for experimental onset and procedures for reprogramming immortalized DIDO3ΔE16. b Top: images of representative wells of AP-positive colonies as a pluripotency marker after infection with OKSM retroviruses, from reprogrammed parental DIDO3floxedE16 before Cre deletion and from DIDO3ΔE16 MEF. Bottom: control wells with GFP-retrovirus infection. c Quantification to determine AP-positive colonies; bars represent mean ± SEM. Statistical analysis using two-tailed Student’s t-test showed significant differences **P ≤ 0.01. d In WT cells, DHX9 binds to RNA pol II and DIDO3 at termination sites, RNA pol II terminates RNA transcription, DHX9 resolves R-loops, and RNA pol II dissociates from DNA. e In DIDO3ΔE16 cells, DHX9 function is impaired, R-loops persist, RNA pol II continues reading, DNA damage occurs on the single-stranded DNA, and transcription–replication conflicts provoke cell stress. All these effects abolish differentiation and reprogramming.

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