To verify the ribosome skip capability of the "2A" sequences in mammalian cells, several constructs were cloned: constructs with the T2A sequence and various linkers downstream of it as well as a construct with the P2A sequence. The constructs were sequence verified prior to experimentation. When they were deposited at Addgene, they were also NGS sequenced in their entirety for their QC process. All plasmids passed the QC. The plasmids are available at http://www.addgene.org/browse/article/28196220/. The...

Pre-publication supplementary data from production of a mouse strain.
A Csrp2-T2A-iCreERT2 knock-in mouse.
Csrp2 was selected as a potential marker of Gfap+ Prom1+ stem cells from analysis of the microarray data in Beckervordersandforth et al., 2010 (https://doi.org/10.1016/j.stem.2010.11.017).
Generated with a standard workflow in the Capecchi laboratory: C57BL/6J BAC clone recombineering to generate the targeting vector; electroporation of the G4 B6 x 129 F1 hybrid mouse embryonic stem cells.

The targeting vector was deposited and sequenced in its entirety at...

Eberdingen-Hochdorf. Aiguilles de bronze et de fer. A. Longueur 14,9 cm.

Supplementary Material for article

An R script to analyze the Mizrak et al., 2019 dataset.

Clusters. Selected marker genes RNA in clusters.
Markers for selected subclusters identified by Seurat.

To validate the AverageExpression command results, RT-qPCR measurements from Morizur et al., 2018 (https://doi.org/10.1016/j.stemcr.2018.06.005) were utilized. The trends were very similar, suggesting that normalized scRNA-seq datasets can also reveal the average RNA expression level of cell populations otherwise obtained by FACS sorting and RT-qPCR.

The relative RNA abundance in the lateral wall V-SVZ qNSC's...

An R script to analyze the Mizrak et al., 2019 dataset.

Clusters. Selected marker genes RNA in clusters.
Markers for selected subclusters identified by Seurat.

To validate the AverageExpression command results, RT-qPCR measurements from Morizur et al., 2018 (https://doi.org/10.1016/j.stemcr.2018.06.005) were utilized. The trends were very similar, suggesting that normalized scRNA-seq datasets can also reveal the average RNA expression level of cell populations otherwise obtained by FACS sorting and RT-qPCR.

The relative RNA abundance in the lateral wall V-SVZ qNSC's...

An R script to analyze the Mizrak et al., 2019 dataset.

Clusters. Selected marker genes RNA in clusters.
Markers for selected subclusters identified by Seurat.

To validate the AverageExpression command results, RT-qPCR measurements from Morizur et al., 2018 (https://doi.org/10.1016/j.stemcr.2018.06.005) were utilized. The trends were very similar, suggesting that normalized scRNA-seq datasets can also reveal the average RNA expression level of cell populations otherwise obtained by FACS sorting and RT-qPCR.

The relative RNA abundance in the lateral wall V-SVZ qNSC's...

An R script to analyze the Mizrak et al., 2019 dataset. Mice in this study were 8-10 week old.

Clusters.

Selected marker genes RNA in clusters.
Markers for selected subclusters identified by Seurat.

To validate the AverageExpression command results, RT-qPCR measurements from Morizur et al., 2018 (https://doi.org/10.1016/j.stemcr.2018.06.005) were utilized. The trends were very similar, suggesting that normalized scRNA-seq datasets can also reveal the average RNA expression level of cell populations otherwise obtained by FACS sorting and RT-qPCR.

The relative...

Propagation or diffusion can be defined as the distribution of a phenomenon in a specific environment. This concept has played different roles in a variety of sciences: the development of matter in physics, the extension of thought and cultural model in sociology and anthropology, the spread of disease in medicine, and the development of new innovative technology or product in economics. In this study, the dissemination and extension of Persian equivalents approved by the Academy...

Work done on Id1-LSL-T2A-TagRFP-T-dre-PBD* mice.
Alignment of the Id1-LSL-RFP-drePR allele targeting vector design sequence and actual NGS sequence (https://www.addgene.org/168770/sequences/). There were some differences in the pWS-TK6 backbone. These are not problematic since the plasmid replicated in E. coli, etc. In the vector itself, there were differences in repetitive regions (e.g., ~11200). The disrupting mutation is at ~19800. The 1-bp difference at ~21300 is a silent mutation in PBD*. I think this was an undocumented sequence change...

MEB-FEG pictures of opal coating and colouring metrials on the surface of limestone flake S-ECA-05 coming from the wall of the rock art cave Points Cave (Aiguèze, Gard, France).

Full visualisation of results in 5A all GTEx CNS regions with six target gene sets. The target gene set distribution is shown in blue and the distribution of 10,000 random size-matched gene sets is shown in green. Vertical dotted lines represent the medians of the target gene set and the central median of the 10,000 bootstrap sets (produced using the MitoNuclearCOEXPlorer tool). A. PanelApp adult onset ND. B. PanelApp intracerebral calcification disorders. C. Jansen et...

Pre-publication supplementary data from production of a mouse strain.
A dre / rox reporter with a tandem dimer superfolder GFP knocked in into Hipp11 (also known as Igs2).
Generated with a standard workflow in the Capecchi laboratory: C57BL/6J BAC clone recombineering to generate the targeting vector; electroporation of the G4 B6 x 129 F1 hybrid mouse embryonic stem cells.
The targeting vector was deposited and sequenced in its entirety at Addgene. The NGS sequence is available at https://www.addgene.org/139513/sequences/....

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This is a supplemental data for the forthcoming publication entitled "Proximal tubule cyclophilin D mediates kidney fibrogenesis in obstructive nephropathy". The data demonstrate that cyclophilin D (CypD) in proximal tubule (PT) is a critical factor contributing to kidney fibrosis in unilateral ureteral obstruction (UUO), providing a new paradigm for mitochondria-targeted therapeutics of fibrotic diseases. Supplementary figure 1 shows that PT-CypD knockout suppresses interstitial cell proliferation during UUO.Supplementary figure 2 shows that PT-CypD knockout does not...

Expanded presentation of the flow cytometry data included in the Nam and Capecchi, 2020 paper.
V-SVZ cells were dissociated to single cells, then stained with the goat anti-Lrig1 polyclonal antibody from R & D Systems (https://doi.org/10.6084/m9.figshare.12736304.v1). The primary antibody was visualized with biotinylated donkey anti-goat secondary and streptavidin Alexa 647. In "unstained" samples, the primary antibody staining was omitted; the secondary and streptavidin were stained. Although the signal was still faint, the biotin-streptavidin staining amplified it...

Characterizing N-terminal variants of iCre-ERT2 in the context of a T2A ribosome skip sequence.
Two recent papers, Kats et al., Molecular Cell, 70, 488-501 (2018) and Reinhardt et al., Stem Cell Reports, 14, 1-8 (2020), suggest N-terminal amino acids are important for protein stability (in addition to the previous work in the literature).
This might be a way to modulate the abundance of the second protein. For some proteins, such as the iCre-ERT2 recombinase, that could be...

the spatial-temporal fusion model was applied to simulate the 9 km AP9 data after failure of the active radar. The method makes full use of all the historical The Soil Moisture Active Passive (SMAP) data at 9 km and 36 km data available between April and July 2015. As a result, 8-day composited 9 km SMAP data at the global scale were produced from 2015 to 2020, by downscaling the corresponding 8-day composited P36 data.

Figures of this study

Supplemental Figure S2. Effect of Ca [a], P [b], and K [c] [reported as mean values (mg/L) of each quintile of the distributions] on gelation, curd-firming and syneresis of individual milk samples using model M2 (no fat and casein; contemporary inclusion of minerals).

Boves, Château, US4432, LM2.In situ fragmentation of a heated fishbone.This fishbone has been dropped on a floor made with silty material and limestone sand. Trampling has induced the fragmentation of the bone and the incorporation into the floor of small charcoals. The floor cover a previous refuse layer visible at the bottom (charcoal). Ashes and very fine charcoal are visible on top.