Fig. 1: PIN1 regulates RNF168 chromatin retention. | Nature Communications

Fig. 1: PIN1 regulates RNF168 chromatin retention.

From: PIN1-SUMO2/3 motif suppresses excessive RNF168 chromatin accumulation and ubiquitin signaling to promote IR resistance

Fig. 1

A Representative images of RNF168 foci in control (siNTC) or PIN1-depleted cells (siPIN1) after treatment with ionizing radiation (IR), 2 Gy. Scale bars 10 µm. B Quantification of RNF168 foci intensity from A. Data is mean ± s.e.m, n = 170 cells. Source data are provided as a Source Data file. C Western blot of chromatin and soluble fractions or whole cell lysate (WCL) for RNF168, histone H3, vinculin and PIN1. Control and PIN1 depleted U2OS cells were treated with 10 Gy of IR and collected after indicated time points to prepare chromatin, soluble fraction or WCL (performed once). Source data are provided as a Source Data file. D Representative images of myc-RNF168 foci in control (siNTC) or PIN1-depleted cells (siPIN1). Scale bars 10 µm. The right panel shows the fluorescence intensity profiles along the line for myc and γH2AX. E Quantification of myc-RNF168 foci intensity from (D) Data is mean ± s.e.m, n = 113 cells for siNTC and 131 cells for siPIN1. Source data are provided as a Source Data file. F Representative images of myc-RNF168 foci in control (siNTC) or PIN1-depleted cells (siPIN1) after 2 Gy IR. Scale bars 10 µm. G Quantification of myc-RNF168 foci intensity from (F). Data is mean ± s.e.m, n = 142 cells for siNTC and 136 cells for siPIN1. Source data are provided as a Source Data file. H Western blot of chromatin fraction and whole cell lysate (WCL) for myc, histone H3, PIN1 and tubulin in control (siNTC) or PIN1-depleted cells (siPIN1) after treatment with 10 Gy IR. SE: short exposure, LE: Long exposure (Representative of 2 repeats). Source data are provided as a Source Data file. I Quantification of myc-RNF168 intensity after treatment with various PIN1 inhibitors: Juglone (10 µM, 4 hrs), PiB (25 µM, 24 hrs), ATRA (25 µM, 24 hrs). Data is mean ± s.e.m, n = 82 for control, 100 for Juglone and PiB, and 86 for ATRA. Source data are provided as a Source Data file. J Quantification of myc-RNF168 foci intensity after treatment with PIN1 inhibitors and IR. Cells expressing myc-RNF168 were treated with Juglone (10 µM, 4 hrs), PiB (25 µM, 24 hrs), ATRA (25 µM, 24 hrs) before irradiation (2 Gy IR). Data is mean ± s.e.m, n = 82 for control, 84 for Juglone, 77 for PiB, and 74 for ATRA. Source data are provided as a Source Data file.

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