Fig. 6: N- or C-terminal truncation of NPM1 blocked the major molecular functions of LETN. | Cell Research

Fig. 6: N- or C-terminal truncation of NPM1 blocked the major molecular functions of LETN.

From: Mutual dependency between lncRNA LETN and protein NPM1 in controlling the nucleolar structure and functions sustaining cell proliferation

Fig. 6

a–d Full-length NPM1 (NPM1-F), C-terminus-truncated NPM1 (NPM1-ΔC, panel a, c) or N-terminus-truncated NPM1 (NPM1-ΔN, panel b, d) was reintroduced into the NPM1–/– HUH7 cells. The chromatin of these cells was partially digested with MNase (0.1 U/300 µL) upon overexpression of LETN. The resulted DNA fragments were visualized by gel electrophoresis (a, b). Relative expression levels of pre- and mature rRNAs were measured by RT-qPCR upon overexpression of LETN and normalized to the housekeeping gene beta-actin (c, d). The statistical significance levels were obtained by pair-wise comparisons between EV and LETN-OE (a, b). Data show means ± SD of three biological replicates. e–g SIM images showing the nucleus staining by DAPI (blue), mCherry-labeled full-length NPM1 (e), NPM1-ΔC (f), or NPM1-ΔN (g) (red), and MS2-tagged LETN marked by MS2-GFP (green) in the NPM1–/– HUH7 cells. h The degrees of colocalization between LETN and the NPM1 proteins were quantified by the Pearson’s correlation coefficients (PCC). The maximal value of PCC (1.0) indicates perfect colocalization between two fluorescence signals in a cell, whereas PCC = 0 indicates no colocalization.

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