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. 2021 May 22;49(10):5845–5866. doi: 10.1093/nar/gkab386

Figure 4.

Figure 4.

Effects of SANS depletion on Cajal bodies in HEK293T cells. (A–C) Double immunofluorescence staining for coilin (red) and PRPF31 in siCtrl- and siSANS- or siPRPF6-treated cells (A) and U5-52K (B) SART1 (C) in siCtrl- and siSANS-treated cells (for hSNU114 staining see Supplementary Figure S4D). (D) Quantification of mean intensity ratios of PRPF31, U5-52K and SART1 signals observed in Cajal bodies reveals the accumulation of PRPF31 and SART1 but not U5-52K in Cajal bodies in SANS-depleted cells. (E–G) Analysis of RNA-FISH labelling of U4 (E), U5 (F), and U6 (G), snRNAs (green) in Cajal bodies (anti-coilin, red) in siCtrl- and siSANS-treated cells. (H) Quantification of the mean intensity ratios of accumulation of snRNAs in Cajal bodies. SANS depletion leads to the significant accumulation of U4 and U6 snRNAs but not U5 snRNA in CBs. Blue, DAPI counterstaining for nuclear DNA. ***P values < 0.001, **P values < 0.01, *P values < 0.05.