Fig. 6
From: Loss of SIM2s inhibits RAD51 binding and leads to unresolved replication stress
SIM2s interacts with RAD51 and is necessary for subnuclear RAD51 localization. a MCF7-shSIM2 and MCF7-pSIL cells were treated with DMSO or 0.5 mM HU for 24 h and then fractionated before being probed for RAD51. α-Tubulin and LaminB1 were used as loading controls to verify efficient separation of fractions. b MCF7-shSIM2 and MCF7-pSIL cells were treated with 0.5 mM HU and harvested 2 h later. BRCA1 was immunoprecipitated and lysates were probed for the indicated proteins. c Stabilization and localization of SIM2s was assessed in SUM159-SIM2s-FLAG treated with 0.5 mM HU and fixed at the indicated timepoints before being probed for FLAG. d Quantification of nuclear FLAG from c. e, f Western blot analysis of SIM2 stabilization in e SUM159-SIM2s-FLAG and f MCF7 cells in response to 0.5 mM HU treatment. Arrow indicates predicted molecular weight of SIM2s. Quantification is the ratio of SIM2s to β-actin. g RAD51 was immunoprecipitated in MCF7 cells after 2 h of treatment with HU or DMSO and lysates were probed for the indicated proteins. h Graphical representation showing fork stabilization in cells containing SIM2s and fork collapse with loss of SIM2s