Figure 1. RTF2 is necessary for cellular proliferation and DNA replication.

a, Representative immunoblot of whole cell lysates showing RTF2 levels 72 hrs after transduction of MEFs with Hit & Run pMMP Cre retrovirus⁴¹. b, Representative growth curves in MEFs transduced with pWZL Cre-hygro retrovirus. c, Representative cell cycle profiles from flow cytometry at 72 hrs after Cre. d, Quantification of representative experiment of mean nuclear signal of EdU from EdU-positive cells at 72 hrs after Cre. e, Top: Schematic and representative image of DNA combing. Below: Quantification of representative experiment of replication tract lengths from primary MEFs at 72 hrs after Cre. f, Top: Representative image of inter-origin distances measured within replication clusters. Bottom: Quantification of representative experiment of inter-origin distances (IOD) from primary MEFs at 72 hrs after Cre. g, Quantification of representative experiment of replication tract lengths from SV40-immortalized MEFs stably expressing HA-FLAG empty vector (EV) or Rtf2 at 120 hrs after Cre. Error bars represent standard deviation for b. Experiments were conducted at least three times in biological replicates with consistent results for a-g. Mean shown with red line for d,e,f,g. Experiments were blinded prior to analysis for d,e,f,g. Average fork speeds are listed above each sample for e and g. Outliers removed with ROUT (1%) for e and g. Significance was evaluated by Kruskal-Wallis ANOVA with a Dunn’s post-test.