DOI: 10.5061/dryad.k3r485h
title: Data from: The RNA-binding protein Celf1 post-transcriptionally regulates p27Kip1 and Dnase2b to control fiber cell nuclear degradation in lens development

description: Opacification of the ocular lens, termed cataract, is a common cause of
 blindness. To become transparent, lens fiber cells undergo degradation of
 their organelles, including their nuclei, presenting a fundamental
 question: does signaling/transcription sufficiently explain
 differentiation of cells progressing toward compromised transcriptional
 potential? We report that a conserved RNA-binding protein Celf1
 post-transcriptionally controls key genes to regulate lens fiber cell
 differentiation. Celf1-targeted knockout mice and celf1-knockdown
 zebrafish and Xenopus morphants have severe eye defects/cataract. Celf1
 spatiotemporally down-regulates the cyclin-dependent kinase (Cdk)
 inhibitor p27Kip1 by interacting with its 5’ UTR and mediating translation
 inhibition. Celf1 deficiency causes ectopic up-regulation of p21Cip1.
 Further, Celf1 directly binds to the mRNA of the nuclease Dnase2b to
 maintain its high levels. Together these events are necessary for
 Cdk1-mediated lamin A/C phosphorylation to initiate nuclear envelope
 breakdown and DNA degradation in fiber cells. Moreover, Celf1 controls
 alternative splicing of the membrane-organization factor beta-spectrin and
 regulates F-actin-crosslinking factor Actn2 mRNA levels, thereby
 controlling fiber cell morphology. Thus, we illustrate new Celf1-regulated
 molecular mechanisms in lens development, suggesting that
 post-transcriptional regulatory RNA-binding proteins have evolved
 conserved functions to control vertebrate oculogenesis.
description: RT-qPCR of Dnase2b in
 mouse lens (Fig 3E)RT-qPCR assay data for
 Dnase2b mRNA in WT and Celf1 cKO mouse lens (Fig 3E in Siddam et al. 2018
 PLOS Genetics)Fig 3E
 .xlsxCelf1 CLIP assay on
 mouse lens (Fig 3G Fig 4K Fig 5C)Data for cross-linked
 immunoprecipitation (CLIP) using Celf1 antibody on wild-type mouse lens
 followed by RT-qPCR for specific mRNAs: Dnase2b, p27Kip1, Sptb (Fig 3G,
 Fig 4K, Fig 5C in Siddam et al. 2018 PLOS Genetics)Fig 3G Fig 4K Fig 5C
 .xlsDnase2b 3'
 UTR-luciferase mRNA assay (Fig 3H)RT-qPCR data for
 quantification of luciferase mRNA fused to Dnase2b 3' UTR in cells
 with Celf1 over-expression (Fig 3H in Siddam et al. 2018 PLOS
 Genetics)Fig
 3H.xlsxQuantification of
 p27Kip1 immunofluorescence in mouse lens (Fig 4G)Immunofluorescence
 signal was quantified for p27Kip1 staining in the whole lens area (left)
 and the central fiber cell area (right) of the lens in control and Celf1
 cKO lenses. (Fig. 4G in Siddam et al. 2018 PLOS
 Genetics)Fig
 4G.xlsxp27Kip1 mRNA in Celf1
 cKO and control mouse lens (Fig 4I)RT-qPCR data for
 quantification of p27Kip1 mRNA in control and Celf1 cKO mouse lens (Fig.
 4I in Siddam et al. 2018 PLOS Genetics)Fig
 4I.xlsxLuciferase reporter for
 p27Kip1 5'UTR in Celf1 knockdown (Fig 4M)Data for luciferase
 reporter assay investigating the up-regulation of 5'UTR fused
 reporter in Celf1 knockdown lens cell line (Fig. 4M in Siddam et al. 2018
 PLOS Genetics)Fig 4M
 .xlsxAssay for p21Cip1 mRNA
 in Celf1 cKO mouse lens (Fig 4N)RT-qPCR data for p21Cip1
 mRNA quantification in control and Celf1 cKO mouse lens (Fig. 4N in Siddam
 et al. 2018 PLOS Genetics)Fig
 4N.xlsxQuantification of Actn2
 mRNA in Celf1 cKO mouse lens (Fig 5A)RT-qPCR data for
 quantification of Actn2 mRNA in control and Celf1 cKO mouse lens (Fig. 5A
 in Siddam et al. 2018 PLOS Genetics)Fig
 5A.xlsxQuantification of Sptb
 isoforms in Celf1 cKO mouse lens (Fig 5D)RT-qPCR data for
 quantification of specific Sptb isoforms in control and Celf1 cKO mouse
 lens (Fig. 5D in Siddam et al. 2018 PLOS Genetics)Fig
 5D.xlsxiSyTE analysis of Celf1
 expression in mouse lens (S1 Fig)Microarray data values
 for expression of Celf1 in wild-type mouse lens in three embryonic stages
 compared to whole embryonic body tissue (Supplementary Fig. S1 in Siddam
 et al. 2018 PLOS Genetics)S1
 Fig.xlsxQuantification of Celf1
 mRNA in Celf1 cKO mouse lens (S3 Fig)Data for RT-qPCR
 quantification of Celf1 mRNA in Celf1 cKO mouse lens compared to control
 lens (Supplementary Fig. S3 in Siddam et al. 2018 PLOS
 Genetics)S3
 Fig.xlsxQuantification of ocular
 defects in Xenopus celf1-knockdown embryos (S5 Fig)Data for quantification
 of ocular phenotypes in celf1-knockdown Xenopus laevis embryos subjected
 to morpholino-injection on one side (Supplementary Fig. S5 in Siddam et
 al. 2018 PLOS Genetics)S5
 Fig.xlsxQuantification of lamin
 A/C immunofluorescence in Celf1 cKO mouse lens (S11 Fig)Data for quantification
 of immunofluorescence signal of lamin A/C in control and Celf1 cKO mouse
 lens (Supplementary Fig. S11 in Siddam et al. 2018 PLOS
 Genetics)S11
 Fig.xlsx