Fig. 7: Mediator localized in the region between CBs and HLBs. | Nature Communications

Fig. 7: Mediator localized in the region between CBs and HLBs.

From: The 3′ Pol II pausing at replication-dependent histone genes is regulated by Mediator through Cajal bodies’ association with histone locus bodies

Fig. 7

a–f HeLa cells were fixed with paraformaldehyde and subjected to triple immunofluorescence staining. MED26 (red), MED1 (red), ICE1 (red), ELL (red), LSM11 (red), and FLASH (red) were co-stained with NPAT (green) and Coilin (gray) using specific antibodies. Scale bar, 5 μm. Averaged signals of immunofluorescence centered at NPAT signal and respective line plots for each immunofluorescence experiment are shown on the right. Scale bar, 1 μm. g, h HCT116 cells were stained with anti-MED26 (green), anti-NPAT (green), and anti-Coilin (magenta) antibodies, and the nuclear positions of MED26 (Mediator), NPAT (HLBs), and Coilin (CBs) were observed by stimulated emission depletion (STED) microscopy. Scale bar, 1 μm. Line intensity plots across the particles of NPAT (green), Coilin (magenta), and MED26 (magenta) are shown in the lower panels. i Ratios of ICE1, ELL, LSM11, MED1, MED26, or FLASH colocalization with CBs, HLBs, or both CBs and HLBs. The ratio was calculated using more than 100 nuclei. ICE1, n = 71 particles from 196 nuclei; ELL, n = 122 particles from 240 nuclei; LSM11, n = 161 particles from 242 nuclei; MED1, n = 84 particles from 237 nuclei; MED26, n = 83 particles from 204 nuclei; FLASH, n = 187 particles from 291 nuclei. Source data are provided as a Source Data file. j Model of the relative position of each protein at CBs and HLBs. Mediator was located between HLBs and CBs.

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