Journal: bioRxiv

Article Title: Nonsense, but not frameshift, truncating mutations result in Cyclin D2 stabilisation in an induced pluripotent stem cell model of MPPH

doi: 10.1101/2025.08.29.673076

Figure Lengend Snippet: A - Electropherogram showing both alleles (top and bottom) of CCND2 biallelic frameshift line CCND2 FS-T1 compared to wildtype (middle) causing truncation at positions 266 and 272 on allele 1 and allele 2, respectively. B - illustrates the same but for the second CCND2 biallelic frameshift line CCND2 FS-T2 showing an 11 bp deletion on allele 1 (top) and a different 11 bp deletion on allele 2 (bottom) compared to wildtype (middle), causing CCND2 truncation at position 272 and 274 respectively. C - Sanger sequencing electropherograms indicating with arrows the single base pair change c.814G>T in heterozygous (top) and homozygous (bottom) CCND2 E272*Het and CCND2 E272*Hom CRISPR-Cas9 edited lines compared to wildtype (middle).

Article Snippet: : Rabbit-CCND2 (CST, 1/1,000) and Mouse-B-actin (Ambion, 1/20,000) antibodies were used.

Techniques: Sequencing, CRISPR

Journal: bioRxiv

Article Title: Nonsense, but not frameshift, truncating mutations result in Cyclin D2 stabilisation in an induced pluripotent stem cell model of MPPH

doi: 10.1101/2025.08.29.673076

Figure Lengend Snippet: A - 2D genomic and proteomic structure of CCND2, showing location of MPPH disease causing variants at the C-terminus (bottom) ( Mirzaa et al ., 2014 , Zhao et al ., 2024 ) and likely pathogenic variants from ClinVar in italics (top). Green indicates phosphorylation sites Ser269, Ser271 and Thr280, pink indicates ubiquitination site Lys270. B – Illustration of the CCND2 C- terminus showing the site of truncations, at the gene and protein level, in each CRISPR-Cas9 engineered line in comparison to wildtype (WT). Red hatched regions indicate regions of amino acid sequence changes due to a frameshift mutation. Green and pink highlighted regions signify locations of phosphorylation sites (Ser269, Ser271 and Thr280) and ubiquitination site Lys270, respectively.

Article Snippet: : Rabbit-CCND2 (CST, 1/1,000) and Mouse-B-actin (Ambion, 1/20,000) antibodies were used.

Techniques: Phospho-proteomics, Ubiquitin Proteomics, CRISPR, Comparison, Sequencing, Mutagenesis

Journal: bioRxiv

Article Title: Nonsense, but not frameshift, truncating mutations result in Cyclin D2 stabilisation in an induced pluripotent stem cell model of MPPH

doi: 10.1101/2025.08.29.673076

Figure Lengend Snippet: A - Representative fluorescence images of pluripotency markers on wildtype and mutant iPSC lines, stained for Oct 3/4 (green), SSEA4 (red) and Hoechst (blue). B - Representative western blot of CCND2 in the presence or absence of cycloheximide in CCND2 FS-T1 and CCND2 FS-T2 frameshift lines. C - Representative western blot of CCND2 in the presence or absence of cycloheximide in CCND2 E272*Het and CCND2 E272*Hom nonsense lines.

Article Snippet: : Rabbit-CCND2 (CST, 1/1,000) and Mouse-B-actin (Ambion, 1/20,000) antibodies were used.

Techniques: Fluorescence, Mutagenesis, Staining, Western Blot

Journal: bioRxiv

Article Title: Nonsense, but not frameshift, truncating mutations result in Cyclin D2 stabilisation in an induced pluripotent stem cell model of MPPH

doi: 10.1101/2025.08.29.673076

Figure Lengend Snippet: Sections were stained for PAX6, β-Tubulin, TBR1, NeuN and Hoechst on all CCND2 CRISPR-Cas9 edited lines and compared to wildtype. For each, similar results were obtained from 2 independent batches grown. Scale bar 20um at x10 magnification.

Article Snippet: : Rabbit-CCND2 (CST, 1/1,000) and Mouse-B-actin (Ambion, 1/20,000) antibodies were used.

Techniques: Staining, CRISPR

Journal: iScience

Article Title: circGlis3 promotes β-cell dysfunction by binding to heterogeneous nuclear ribonucleoprotein F and encoding Glis3-348aa protein

doi: 10.1016/j.isci.2023.108680

Figure Lengend Snippet:

Article Snippet: Rabbit monoclonal anti-CCND2 , Cell Signaling Technology , Cat# 3741; RRID: AB_2070685.

Techniques: Virus, Recombinant, Modification, Lysis, Transfection, Enzyme-linked Immunosorbent Assay, Bicinchoninic Acid Protein Assay, CCK-8 Assay, Imaging, TUNEL Assay, Apoptosis Assay, In Situ Hybridization, Immunoprecipitation, Mass Spectrometry, Western Blot, shRNA, Plasmid Preparation, Software