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Addgene inc pme sema7a
Pme Sema7a, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pme+mkate2/p3E-mKate2-myc+no-pA+(Plasmid+%2380812)/pmc11318972-287-17-22
Average 93 stars, based on 1 article reviews

pme sema7a - by Bioz Stars, 2026-10

93/100 stars

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Plasmid Preparation:

Article Title: Mechanism-Based Personalized Medicine for Cystic Fibrosis by Suppressing Pseudo Exon Inclusion.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER pSS21_ CFTRWT-GFP (CFTR CDS+truncated IVS22 (WT) with N-terminal mKate2-GP2A and C-terminal HiBiTHA-AcGFP) This paper N/A pSS22_ CFTRC>T-GFP (CFTR CDS+truncated IVS22 (3849+10kb C>T) with N-terminal mKate2-GP2A and C-terminal HiBiT-HA-AcGFP) This paper N/A pSS23_ CFTRF508d-GFP (CFTR CDS (DF508 ) with N-terminal mKate2-GP2A and C-terminal AcGFP) This paper N/A CFTR WT_HiBiT Promega N/A (Gift from Promega) CFTR_delF508_HiBiT Promega N/A (Gift from Promega) pME-mKate2 no stop vector Fowler et al., 2016 Addgene plasmid # 80806 (pME-mKate2 no stop) pcDNA3.1 EYFP H148Q/I152L Galietta et al., 2001 Addgene plasmid # 25872 (pcDNA3.1 EYFP H148Q/I152L) Software and Algorithms GraphPad Prism Version 8 GraphPad Software, San Diego, CA, USA www.graphpad.com Hybrid cell count BZ-H3C software Keyence, Osaka, Japan https://www.keyence.com/ Image Lab software Bio-Rad Laboratories, Inc., Hercules, CA, USA https://www.bio-rad.com/ Cell Chemical Biology 27, 1–11.e1–e6, December 17, 2020 e2 system, the normal-type splicing (skipping of the pseudo exon) leads to the expression of a GFP-fusion protein. ..

Software:

Cell Characterization:

Expressing:

Article Title: Mechanism-Based Personalized Medicine for Cystic Fibrosis by Suppressing Pseudo Exon Inclusion.
Article Snippet: .. The F508del-CFTR expression vector and the HiBiT sequence was provided by Promega Corporation (Madison, WI, USA), andmKate2was subcloned from pME-mKate2 no stop vector by Philip Washbourne (Addgene #80806) (Fowler et al., 2016). ..

Sequencing:

Article Title: Distinct progenitor populations mediate regeneration in the zebrafish lateral line
Article Snippet: The myo6b:mKate2 construct was generated via the Gateway Tol2 system (Invitrogen). .. A pME-mKate2 (the mKate2 sequence being cloned from pMTB-Multibow-mfR, Addgene #60991) construct was generated via BP Recombination, and then a pDEST-myo6b:mKate2 construct was generated via LR recombination of p5E-myo6b, pME-mKate2, p3E-pA, and pDEST-iTol2-pA2 vectors. ..

Clone Assay:

Construct:

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Image Search Results

(A) LEFT Depiction of mechanical compression assay where animal is mounted dorsally on inverted spinning disk confocal with a dextran loaded microneedle mounted above the animal. RIGHT image of mechanical compression assay apparatus. (B) Depiction of mechanical compression assay with needle placing force on DRG. (C) Quantification of the percent of DRG responding to mechanical force in animals expressing Tg(sox10 : gal4+myl7); Tg(uas : GCaMP6s) (labeled sox10) or expressing Tg(neurod : gal4+myl7); Tg(uas : GCaMP6s) (labeled neurod) and treated with either DMSO or GsMTx4 at both 2 and 3 dpf (sox10 2 dpf DMSO: n = 7 animals, 7 DRG, sox10 2 dpf GsMTx4: n = 5 animals, 5 DRG neurod 2 dpf DMSO: n = 5 animals 5 DRG neurod 2 dpf GsMTx4: n = 4 animals, 4 DRG sox10 3 dpf DMSO: n = 11 animals, 11 DRG sox10 3 dpf GsMTx4: n = 4 animals, 4 DRG neurod 3 dpf DMSO: n = 9 animals, 9 DRG neurod 3 dpf GsMTx4: n = 4 animals, 4 DRG). (D) Confocal image taken of the mechanical compression assay in 2 dpf animal expressing Tg(sox10 : gal4+myl7); Tg(uas : GCaMP6s) . Images show an inactive time point, a time point with tissue compression, and an active time point in response to tissue compression. Inactive and active DRG marked with an arrow. (E) Quantification of the change in integrated density of fluorescence during each phase of mechanical compression assay of a DRG in a 3 dpf animal treated with either DMSO or GsMTx4. Change in integrated density of fluorescence is scored as time point subtracting the initial time point divided by time point ( Δf / f ). (F) Quantification of the average change in fluorescence during the phases of mechanical compression following treatment with either 2% DMSO or 1 μM GsMTx4 for 30 min (Resting, Compression, and Decompression DMSO: n = 9 DRG, 9 animals, Resting, Compression, and Decompression GsMTx4: n = 4 DRG, 4 animals). (G) Confocal z-projection of peripheral DRG axon in an animal expressing Tg(ngn1 : GFP) at 2 and 3 dpf. Arrow notes the end processes of the peripheral axon. Arrowhead denotes peripheral axons from Rohon beard neurons. (H) Average distance (μM) of DRG displacement needed to elicit a response ( n = 16 animals, 16 DRG). (I) Confocal images of RNAscope- piezo1 and Immunohistochemistry-GFP in Tg(sox10 : meGFP) animals. GFP is shown in magenta and piezo1 is shown in cyan. Arrowheads indicate piezo1 puncta. Arrows indicate autofluorescence. (J) Quantification of DRG at 3 dpf with piezo1 puncta and without piezo1 puncta ( n = 8 animals, 24 DRG). (K) Confocal images of 3 dpf animals expressing Tg(sox10 : gal4+myl7); Tg(uas : GCaMP6s) . Red colors indicate a higher intensity of fluorescence and blue colors indicate a lower intensity of fluorescence. Images depicted are of animals either treated with 2% DMSO, 40 μM Jedi2, or 1 μM GsMTx4. Arrows note active cells. (L) Line graphs of z score of integrated density of fluorescence for a 1-h time period in 3 dpf animals expressing Tg(sox10 : gal4+myl7); Tg(uas : GCaMP6s) that were treated with either 2% DMSO, 40 μM Jedi2, or 1 μM GsMTx4. A z score greater than 2.58 indicates an active Ca 2+ event. Red scale bar shows a z score of 2.58. (M) Heatmaps of the z score of individual sox10 + cells from animals in G and H during a 1-h period of Ca 2+ imaging. Yellow notes a high z score (2.58 or greater) (DMSO: n = 8 animals, 17 DRG, 52 cells, Jedi2: n = 7 animals, 18 DRG, 79 cells, GsMTx4: n = 6 animals, 16 DRG, 44 cells). (N) Quantification of the average number of Ca 2+ events per sox10 + cell in animals treated with either 2% DMSO, 1 μM GsMTx4, 100 μM Yoda1, or 40 μM Jedi2 (DMSO: n = 8 animals, 17 DRG, 52 cells, Jedi2: n = 7 animals, 18 DRG, 79 cells, GsMTx4: n = 6 animals, 16 DRG, 44 cells, Yoda1: n = 4 animals, 9 DRG, 35 cells). (O) Quantification of the average number of Ca 2+ events per neurod + cell in 3 dpf animals expressing Tg(neurod : gal4+myl7); Tg(uas : GCaMP6s) that were treated with either 2% DMSO, 1 μM GsMTx4, 40 μM Jedi2, or 100 μM Yoda1 (DMSO: n = 10 animals, 24 DRG, 33 cells, Jedi2: n = 5 animals, 18 DRG, 35 cells, GsMTx4: n = 4 animals, 8 DRG, 8 cells, Yoda1: 4 animals, 7 DRG, 8 cells). (P) Quantification of the average number of Ca 2+ events per sox10 + cell following genetic manipulation via injection of uas : cas9mkate-u6 : piezo1gRNA or uas : cas9mkate-u6 : emptygRNA into animals expressing Tg(sox10 : gal4+myl7); Tg(uas : GCaMP6s) at 3 dpf. Additionally, a group of Tg(sox10 : gal4+myl7); Tg(uas : GCaMP6s) injected with uas : cas9mkate-u6 : piezo1gRNA and treated with 40 μM Jedi2 treatment was also included in the experiment (u6:emptygRNA: n = 6 animals, 16 DRG, 40 cells, u6:piezo1gRNA: n = 4 animals, 13 DRG, 57 cells, u6:piezo1gRNA+Jedi2: n = 4 animals, 4 DRG, 7 cells). Scale bar is 10 μM (D, G, I, K). Statistical tests: unpaired t test (H, N, O), Fisher’s exact (C), multiple unpaired t tests (F). The data underlying this figure can be found in . dpf, days post fertilization; DRG, dorsal root ganglia.

Journal: PLOS Biology

Article Title: Piezo1-mediated spontaneous calcium transients in satellite glia impact dorsal root ganglia development

doi: 10.1371/journal.pbio.3002319

Figure Lengend Snippet: (A) LEFT Depiction of mechanical compression assay where animal is mounted dorsally on inverted spinning disk confocal with a dextran loaded microneedle mounted above the animal. RIGHT image of mechanical compression assay apparatus. (B) Depiction of mechanical compression assay with needle placing force on DRG. (C) Quantification of the percent of DRG responding to mechanical force in animals expressing Tg(sox10 : gal4+myl7); Tg(uas : GCaMP6s) (labeled sox10) or expressing Tg(neurod : gal4+myl7); Tg(uas : GCaMP6s) (labeled neurod) and treated with either DMSO or GsMTx4 at both 2 and 3 dpf (sox10 2 dpf DMSO: n = 7 animals, 7 DRG, sox10 2 dpf GsMTx4: n = 5 animals, 5 DRG neurod 2 dpf DMSO: n = 5 animals 5 DRG neurod 2 dpf GsMTx4: n = 4 animals, 4 DRG sox10 3 dpf DMSO: n = 11 animals, 11 DRG sox10 3 dpf GsMTx4: n = 4 animals, 4 DRG neurod 3 dpf DMSO: n = 9 animals, 9 DRG neurod 3 dpf GsMTx4: n = 4 animals, 4 DRG). (D) Confocal image taken of the mechanical compression assay in 2 dpf animal expressing Tg(sox10 : gal4+myl7); Tg(uas : GCaMP6s) . Images show an inactive time point, a time point with tissue compression, and an active time point in response to tissue compression. Inactive and active DRG marked with an arrow. (E) Quantification of the change in integrated density of fluorescence during each phase of mechanical compression assay of a DRG in a 3 dpf animal treated with either DMSO or GsMTx4. Change in integrated density of fluorescence is scored as time point subtracting the initial time point divided by time point ( Δf / f ). (F) Quantification of the average change in fluorescence during the phases of mechanical compression following treatment with either 2% DMSO or 1 μM GsMTx4 for 30 min (Resting, Compression, and Decompression DMSO: n = 9 DRG, 9 animals, Resting, Compression, and Decompression GsMTx4: n = 4 DRG, 4 animals). (G) Confocal z-projection of peripheral DRG axon in an animal expressing Tg(ngn1 : GFP) at 2 and 3 dpf. Arrow notes the end processes of the peripheral axon. Arrowhead denotes peripheral axons from Rohon beard neurons. (H) Average distance (μM) of DRG displacement needed to elicit a response ( n = 16 animals, 16 DRG). (I) Confocal images of RNAscope- piezo1 and Immunohistochemistry-GFP in Tg(sox10 : meGFP) animals. GFP is shown in magenta and piezo1 is shown in cyan. Arrowheads indicate piezo1 puncta. Arrows indicate autofluorescence. (J) Quantification of DRG at 3 dpf with piezo1 puncta and without piezo1 puncta ( n = 8 animals, 24 DRG). (K) Confocal images of 3 dpf animals expressing Tg(sox10 : gal4+myl7); Tg(uas : GCaMP6s) . Red colors indicate a higher intensity of fluorescence and blue colors indicate a lower intensity of fluorescence. Images depicted are of animals either treated with 2% DMSO, 40 μM Jedi2, or 1 μM GsMTx4. Arrows note active cells. (L) Line graphs of z score of integrated density of fluorescence for a 1-h time period in 3 dpf animals expressing Tg(sox10 : gal4+myl7); Tg(uas : GCaMP6s) that were treated with either 2% DMSO, 40 μM Jedi2, or 1 μM GsMTx4. A z score greater than 2.58 indicates an active Ca 2+ event. Red scale bar shows a z score of 2.58. (M) Heatmaps of the z score of individual sox10 + cells from animals in G and H during a 1-h period of Ca 2+ imaging. Yellow notes a high z score (2.58 or greater) (DMSO: n = 8 animals, 17 DRG, 52 cells, Jedi2: n = 7 animals, 18 DRG, 79 cells, GsMTx4: n = 6 animals, 16 DRG, 44 cells). (N) Quantification of the average number of Ca 2+ events per sox10 + cell in animals treated with either 2% DMSO, 1 μM GsMTx4, 100 μM Yoda1, or 40 μM Jedi2 (DMSO: n = 8 animals, 17 DRG, 52 cells, Jedi2: n = 7 animals, 18 DRG, 79 cells, GsMTx4: n = 6 animals, 16 DRG, 44 cells, Yoda1: n = 4 animals, 9 DRG, 35 cells). (O) Quantification of the average number of Ca 2+ events per neurod + cell in 3 dpf animals expressing Tg(neurod : gal4+myl7); Tg(uas : GCaMP6s) that were treated with either 2% DMSO, 1 μM GsMTx4, 40 μM Jedi2, or 100 μM Yoda1 (DMSO: n = 10 animals, 24 DRG, 33 cells, Jedi2: n = 5 animals, 18 DRG, 35 cells, GsMTx4: n = 4 animals, 8 DRG, 8 cells, Yoda1: 4 animals, 7 DRG, 8 cells). (P) Quantification of the average number of Ca 2+ events per sox10 + cell following genetic manipulation via injection of uas : cas9mkate-u6 : piezo1gRNA or uas : cas9mkate-u6 : emptygRNA into animals expressing Tg(sox10 : gal4+myl7); Tg(uas : GCaMP6s) at 3 dpf. Additionally, a group of Tg(sox10 : gal4+myl7); Tg(uas : GCaMP6s) injected with uas : cas9mkate-u6 : piezo1gRNA and treated with 40 μM Jedi2 treatment was also included in the experiment (u6:emptygRNA: n = 6 animals, 16 DRG, 40 cells, u6:piezo1gRNA: n = 4 animals, 13 DRG, 57 cells, u6:piezo1gRNA+Jedi2: n = 4 animals, 4 DRG, 7 cells). Scale bar is 10 μM (D, G, I, K). Statistical tests: unpaired t test (H, N, O), Fisher’s exact (C), multiple unpaired t tests (F). The data underlying this figure can be found in . dpf, days post fertilization; DRG, dorsal root ganglia.

Article Snippet: To create uas : cas9mkate-u6 : piezo1gRNA , we recombined a p5E-UAS (BseRI cleavage site removed), pME-cas9mkate (Addgene 109547), and p3E-pA constructs into a pDestTol2CG2 containing a U6 promoter sequence flanked with BseRI cleavage sites [ , ].

Techniques: Expressing, Labeling, Fluorescence, RNAscope, Immunohistochemistry, Imaging, Injection

(A) Confocal z-projection of DRG in 3 dpf animal expressing Tg(sox10 : gal4+myl7); Tg(uas : GCaMP6s) and treated with either 2% DMSO or 40 μM Jedi2. Individual cells are traced for ROIs and labeled with a number. (B) Line graphs of the z score of the integrated density of fluorescence over a 1 h period. Each numbered line graph corresponds to a numbered ROI. Red scale bar represents a z score of 2.58. Blue arrowheads note simultaneously active time points. Red arrowheads note isolated active time points. (C) Quantification of the average number of isolated Ca 2+ transient events per sox10 + cell in 3 dpf animals expressing Tg(sox10 : gal4+myl7); Tg(uas : GCaMP6s) that were treated with either 2% DMSO, 1 μM GsMTx4, 100 μM Yoda1, or 40 μM Jedi2 (DMSO: 4 animals, 5 DRG, 24 cells, GsMTx4: n = 3 animals, 5 DRG, 23 cells, Jedi2: n = 7 animals, 18 DRG, 59 cells, Yoda1: n = 3 animals, 6 DRG, 28 cells). (D) Quantification of the average number of simultaneous Ca 2+ transient events per sox10 + cell in 3 dpf animals expressing Tg(sox10 : gal4+myl7); Tg(uas : GCaMP6s) that were treated with either 2% DMSO, 1 μM GsMTx4, 100 μM Yoda1, or 40 μM Jedi2 (DMSO: n = 4 animals, 5 DRG, 24 cells, GsMTx4: n = 3 animals, 5 DRG, 23 cells, Jedi2: n = 7 animals, 18 DRG, 59 cells, Yoda1: n = 3 animals, 6 DRG, 28 cells). (E) Quantification of the average number of isolated Ca 2+ transient events following genetic manipulation via CRISPR/Cas9 targeting piezo1 or empty gRNA cassette in 3 dpf animals expressing Tg(sox10 : gal4+myl7); Tg(uas : GCaMP6s) uas : cas9mkate-u6 : piezo1gRNA or uas : cas9mkate-u6 : emptygRNA (u6:emptygRNA: n = 6 animals, 14 DRG, 38 cells, u6:piezo1gRNA: n = 4 animals, 13 DRG, 57 cells). (F) Quantification of the average number of simultaneous Ca 2+ transient events following genetic manipulation via CRISPR/Cas9 targeting piezo1 or empty gRNA cassette in 3 dpf animals expressing Tg(sox10 : gal4+myl7); Tg(uas : GCaMP6s) uas : cas9mkate-u6 : piezo1gRNA or uas : cas9mkate-u6 : emptrygRNA (u6:emptygRNA: n = 6 animals, 14 DRG, 38 cells, u6:piezo1gRNA: n = 4 animals, 13 DRG, 57 cells). (G) Quantification of the average number of microdomains per DRG in 3 dpf animals expressing Tg(sox10 : gal4+myl7); uas : GCaMP6s-caax that were treated with either 2% DMSO, 1 μM GsMTx4, or 40 μM Jedi2 for 30 min prior to imaging (DMSO: n = 7 animals, 7 DRG, GsMTx4: n = 7 animals, 7 DRG, Jedi2: n = 6 animals, 8 DRG). (H) Quantification of the average duration of microdomains in 3 dpf animals expressing Tg(sox10 : gal4+myl7); uas : GCaMP6s-caax that were treated with either 2% DMSO or 40 μM Jedi2 for 30 min prior to imaging (DMSO: n = 8 animals, 8 DRG, Jedi2: n = 5 animals, 7 DRG). Scale bar is 10 μM (A). Statistical tests: unpaired t test (C, D, E, F, H), one-way ANOVA followed and represented by post hoc Dunnett test (G). The data underlying this figure can be found in . dpf, days post fertilization; DRG, dorsal root ganglia; ROI, regions of interest.

Journal: PLOS Biology

Article Title: Piezo1-mediated spontaneous calcium transients in satellite glia impact dorsal root ganglia development

doi: 10.1371/journal.pbio.3002319

Figure Lengend Snippet: (A) Confocal z-projection of DRG in 3 dpf animal expressing Tg(sox10 : gal4+myl7); Tg(uas : GCaMP6s) and treated with either 2% DMSO or 40 μM Jedi2. Individual cells are traced for ROIs and labeled with a number. (B) Line graphs of the z score of the integrated density of fluorescence over a 1 h period. Each numbered line graph corresponds to a numbered ROI. Red scale bar represents a z score of 2.58. Blue arrowheads note simultaneously active time points. Red arrowheads note isolated active time points. (C) Quantification of the average number of isolated Ca 2+ transient events per sox10 + cell in 3 dpf animals expressing Tg(sox10 : gal4+myl7); Tg(uas : GCaMP6s) that were treated with either 2% DMSO, 1 μM GsMTx4, 100 μM Yoda1, or 40 μM Jedi2 (DMSO: 4 animals, 5 DRG, 24 cells, GsMTx4: n = 3 animals, 5 DRG, 23 cells, Jedi2: n = 7 animals, 18 DRG, 59 cells, Yoda1: n = 3 animals, 6 DRG, 28 cells). (D) Quantification of the average number of simultaneous Ca 2+ transient events per sox10 + cell in 3 dpf animals expressing Tg(sox10 : gal4+myl7); Tg(uas : GCaMP6s) that were treated with either 2% DMSO, 1 μM GsMTx4, 100 μM Yoda1, or 40 μM Jedi2 (DMSO: n = 4 animals, 5 DRG, 24 cells, GsMTx4: n = 3 animals, 5 DRG, 23 cells, Jedi2: n = 7 animals, 18 DRG, 59 cells, Yoda1: n = 3 animals, 6 DRG, 28 cells). (E) Quantification of the average number of isolated Ca 2+ transient events following genetic manipulation via CRISPR/Cas9 targeting piezo1 or empty gRNA cassette in 3 dpf animals expressing Tg(sox10 : gal4+myl7); Tg(uas : GCaMP6s) uas : cas9mkate-u6 : piezo1gRNA or uas : cas9mkate-u6 : emptygRNA (u6:emptygRNA: n = 6 animals, 14 DRG, 38 cells, u6:piezo1gRNA: n = 4 animals, 13 DRG, 57 cells). (F) Quantification of the average number of simultaneous Ca 2+ transient events following genetic manipulation via CRISPR/Cas9 targeting piezo1 or empty gRNA cassette in 3 dpf animals expressing Tg(sox10 : gal4+myl7); Tg(uas : GCaMP6s) uas : cas9mkate-u6 : piezo1gRNA or uas : cas9mkate-u6 : emptrygRNA (u6:emptygRNA: n = 6 animals, 14 DRG, 38 cells, u6:piezo1gRNA: n = 4 animals, 13 DRG, 57 cells). (G) Quantification of the average number of microdomains per DRG in 3 dpf animals expressing Tg(sox10 : gal4+myl7); uas : GCaMP6s-caax that were treated with either 2% DMSO, 1 μM GsMTx4, or 40 μM Jedi2 for 30 min prior to imaging (DMSO: n = 7 animals, 7 DRG, GsMTx4: n = 7 animals, 7 DRG, Jedi2: n = 6 animals, 8 DRG). (H) Quantification of the average duration of microdomains in 3 dpf animals expressing Tg(sox10 : gal4+myl7); uas : GCaMP6s-caax that were treated with either 2% DMSO or 40 μM Jedi2 for 30 min prior to imaging (DMSO: n = 8 animals, 8 DRG, Jedi2: n = 5 animals, 7 DRG). Scale bar is 10 μM (A). Statistical tests: unpaired t test (C, D, E, F, H), one-way ANOVA followed and represented by post hoc Dunnett test (G). The data underlying this figure can be found in . dpf, days post fertilization; DRG, dorsal root ganglia; ROI, regions of interest.

Techniques: Expressing, Labeling, Fluorescence, Isolation, CRISPR, Imaging

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pme sema7a (Addgene inc)

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