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. 2023 Apr 10;222(5):e202106022. doi: 10.1083/jcb.202106022

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© 2023 Moore et al.

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Figure S3. — RFWD3 promotes the reversal of stalled replication forks. (A and B) Representative images of normal (A) and reversed (B) replication forks detected by electron microscopy (EM) in control or RFWD3-depleted U2OS upon HU treatment (2 mM for 5 h). Quantitation of reversed forks is provided in Fig. 5 E and Fig. S3 C. (C) Percentage of reversed replication forks identified by EM in the three independent experiments quantitated in Fig. 5 E. The number of replication intermediates is indicated in parentheses. Immunoblot figures showing the efficiency of RFWD3 depletion for each experiment are specified. Decrease in reversed forks correlates with the efficiency of RFWD3 depletion, which is weaker for the third experiment. Reversed fork values for the siFF and siRFWD3 conditions are effectively paired, as the first experiment was performed with a less potent preparation of HU (siFF vs. siRFWD3, P < 0.01 by paired t test, P < 0.05 by unpaired t test).