Embroidery software is software that helps users create embroidery designs. While a large majority of embroidery software is specific to machine embroidery, there is also software available for use with hand embroidery, such as cross-stitch..
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Download Fusion 360 as a 30-day free trial here. Existing subscribers can also access Fusion 360 in their Autodesk account.\n"}]},"@type":"Question","name":"How can I download Fusion 360 for free?","acceptedAnswer":["@type":"Answer","text":"Fusion 360 can be downloaded as a free commercial trial for 30 days, or as a personal use subscription with limited functionality.\n"],"@type":"Question","name":"Is Fusion 360 still free for hobbyists or for personal use?","acceptedAnswer":["@type":"Answer","text":"Fusion 360 is available for free personal use for individuals who are doing home-based, non-commercial design, manufacturing, and fabrication projects.\n"],"@type":"Question","name":"Is Fusion 360 free for students?","acceptedAnswer":["@type":"Answer","text":"Fusion 360 is free cloud-based 3D CAD, CAM, CAE and PCB software for qualifying students as a 1-year subscription. Download Fusion 360 for students\n"],"@type":"Question","name":"Is Fusion 360 free for schools?","acceptedAnswer":["@type":"Answer","text":"Fusion 360 is free software for educators, academic institutions, and students as a 1-year subscription. Download Fusion 360 for schools\n"]],"@type":"FAQPage","@context":" "} Autodesk Company overview Careers Investor relations Newsroom Diversity and belonging
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iMovie for iOS and iMovie for macOS are designed to work together. You can start cutting a project on your iPhone, then use AirDrop or iCloud Drive to wirelessly transfer it to your iPad. You can also send a project from your iPhone or iPad to your Mac for finishing touches like color correction and animated maps. And you can even open iMovie projects in Final Cut Pro to take advantage of professional editing tools. Time to take a bow.
We designed a convergent primer and a divergent primer for the loop-forming sites of circALG1. Agarose gel electrophoresis showed that the convergent primer amplified bands in both cDNA and gDNA, whereas the divergent primer only amplified bands in cDNA. Further cDNA sequencing revealed the presence of circularization sites (Fig. 2A), indicating the circularity of circALG1. Compared with the parental gene ALG1 mRNA, circALG1 showed resistance to RNase R digestion (Fig. 2B). After actinomycin treatment, the degradation rate of circALG1 in cells was significantly slower than that of ALG1 mRNA (Fig. 2C), which indicated that circALG1 was stable. The nucleus-cytoplasm separation assay demonstrated that circALG1 was mainly localized in the cytoplasm (Fig. 2D). Similar results were obtained from circALG1 FISH (Fig. 2E).
We constructed stably transfected HCT116 cells that overexpressed circALG1, and the overexpression efficiency is shown in Supplementary Fig. 1C. We designed 3 siRNAs for circALG1 interference, and the knockdown efficiency is shown in Supplementary Fig. 1D. The si-3 sequence was used to construct a lentivirus that interfered with circALG1 expression, and a stably transfected cell line was generated. The Transwell assay results demonstrated that the migration and invasion abilities of HCT116 cells were enhanced after circALG1 overexpression (Fig. 3A), whereas the migration and invasion abilities of SW480 cells were weakened after the silencing of circALG1 expression (Fig. 3B).
Then, we tested whether the parental linear ALG1 could affect the function of circALG1. qRT-PCR showed that the overexpression and interference of circALG1 had no effect on the expression of linear ALG1 mRNA (Supplementary Fig. 2A). We also constructed an ALG1 overexpression plasmid and transfected it into HCT116 and SW480 cells. The overexpression efficiency is shown in Supplementary Fig. 2B. We designed 3 siRNAs for ALG1 interference, and the knockdown efficiency is shown in Supplementary Fig. 2B. Transwell assays showed that ALG1 was not associated with any significant change in migration or invasion of HCT116 and SW480 cells (Supplementary Fig. 2C). These results showed that parental linear ALG1 did not contribute to the function of circALG1.
To study the interaction between these 5 miRNAs and circALG1, we conducted the following experiment. The SV40-firefly_Luciferase-MCS plasmid containing the circALG1 wild-type sequence was constructed and cotransfected with miRNA mimics (Supplementary Fig. 4A) and a Renilla luciferase plasmid. A decrease in fluorescence intensity was observed in the miR-342-5p group (Fig. 5A), which suggested that miR-342-5p might bind to circALG1. We then assessed the expression level of miR-342-5p in CRC and found that miR-342-5p expression was low in CRC tissues (Supplementary Fig. 4B). Based on the predicted binding sites of circALG1 and miR-342-5p, we designed an SV40-firefly_Luciferase-MCS plasmid containing the circALG1 mutant sequence (Fig. 5B, left). The dual-luciferase reporter assay results indicated that after transfection of the firefly luciferase plasmid containing the circALG1 wild-type sequence, miR-342-5p mimics significantly reduced the fluorescence intensity, whereas the miR-342-5p inhibitor enhanced the fluorescence intensity. No difference was detected with the plasmid containing the circALG1 mutant sequence (Fig. 5B, right, Supplementary Fig. 4C), which indicated that miR-342-5p specifically bound to circALG1.
We then investigated the specific mechanism through which m6A modification affects the binding of circALG1 to miR-342-5p. The literature suggests that the function of circRNA m6A modification often requires the involvement of m6A modification-related proteins [24,25,26]. MS detection of the proteins pulled down in the circALG1 RAP assay showed that YTHDF1 enrichment was highest among the m6A modification-related proteins; therefore, this protein was preliminarily selected as the focus of study. The presence of YTHDF1 was also observed among the pulled-down proteins (Fig. 7C). Additionally, we detected circALG1 enrichment in the YTHDF1 RIP assay (Fig. 7D). When the circALG1 m6A modification site was mutated, the YTHDF1 RIP assay results indicated that the enriched circALG1 was significantly reduced, which suggested that the binding of circALG1 to YTHDF1 was dependent on m6A modification (Fig. 7E). To further study the effect of YTHDF1 on circALG1 function, we designed siRNA targeting YTHDF1, and the siRNA interference efficiency is shown in Supplementary Fig. 6A. The results showed that the downregulation of YTHDF1 expression reduced the binding ability of circALG1 to miR-342-5p (Fig. 7F) and decreased the expression level of the target gene, i.e., PGF, in the circALG1/miR-342-5p signalling axis (Fig. 7G) through the specific mechanism of YTHDF1 enhancing the stability of circALG1 (Fig. 7H).
Changwei Lin and Min Ma, Experimental design, Cell experiment, Data analysis. Yi Zhang, Liang Li and Fei Long, Experimental design, Cell experiment. Canbin Xie, Hua Xiao and Teng Liu, Molecular biology experiment. Buning Tian, Kaiyan Yang and Yihang Guo, Animal experiment. Miao Chen, Jin Chou and Ni Gong, Data analysis. Xiaorong Li and Gui Hu, Supervision, Investigation, Methodology, Writing - original draft, Writing. The author(s) read and approved the final manuscript.
LYL, LBG, and RZ contributed to the study design. LYL and ML contributed to data analysis, interpretation of data and manuscript writing. LYL, YJG, XHW, and YXY performed the experiments, collected and organized clinical samples, and prepared the figures. All authors read and approved the final manuscript.
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We have packed this digtizing software with so many time-saving features that will help you create more in less time. Imagine the capability to create your own fonts, stitches and designs - just once - to use many times on hats, pillows, shirts, and other fabrics. Reduce the amount of time spent changing threads by using the Intelligent Colour Sort feature, which will stitch those areas that share one colour in groups.
Having the ability to choose your own built-in fonts and designs in less time will help you start your projects sooner. Using the font filter will help you narrow down your choices from the 130 fonts built into PE-DESIGN 11. Choosing from over 1,000 built-in designs is now easier with the design preview that lets you see your choice before you stitch.
Y.G. and F.G.W. conceived the research and designed the experiments. Y.G., S.Z.W., Xinping Z., S.Y.W., and Y.L. performed the experiments. Y.G., H.R.J., Y.X.Z., Xiaodong Z., G.G., Y.W.J., C.L., X.C., and F.G.W. analyzed the data. Y.G. and F.G.W. wrote the manuscript. F.G.W. supervised the project. All authors read and approved the manuscript. 2ff7e9595c
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