Fig. 5: lncREST interacts with the replication fork and replication factors. | Nature Communications

Fig. 5: lncREST interacts with the replication fork and replication factors.

From: The chromatin-associated lncREST ensures effective replication stress response by promoting the assembly of fork signaling factors

Fig. 5

a Schematic of the adapted iPOND protocol applied to pulldown the RNAs associated to the replication fork. b Western blot analysis of the fractions obtained from the different pulses with markers of nascent chromatin (PCNA), mature chromatin (H3) and DNA repair (RAD51). A representative experiment of at least three repetitions. Source data are provided as a Source Data file. c qRT-PCR showing the amount of lncREST and MALAT1 relative to the EdU sample (nascent chromatin) in NT (left) and HU treated (right) samples. Each sample was normalized to its input. **p = 0.0052 EdU vs no click NT, **p = 0.0051 EdU+Thy vs EdU, **p = 0.0021 EdU vs no click HU, ***p = 0.0006 EdU+Thy vs EdU HU (mean ± SD, two-tailed unpaired t-test. n = 3). d qRT-PCR showing the efficiency of lncREST pulldown by specific biotinylated probes with respect to LacZ probes. The lncRNA CONCR and HPRT were used as negative controls. **p = 0.0013 lncREST enrichment in pulldown; *p < 0.001 (mean ± SD, two-tailed unpaired t-test. n = 3). e Pie chart showing the percentage of proteins identified as lncREST interactors in the different cellular fractions. f Dot plot indicating the number of peptides detected of each of the proteins identified as lncREST interactors in two independent experiments. For b–f, source data are provided as a Source Data file.

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