standards gal 2 dag 1 2 diacyl 3 o α d galactosyl1 6 β d galactosyl sn glycerol avanti polar lipids 840524p  (Croda International Plc)

Journal: Skeletal Muscle

Article Title: Functional and structural pathologies in skeletal muscle of a rat model of Duchenne muscular dystrophy

doi: 10.1186/s13395-026-00419-4

Figure Lengend Snippet: CRISPR/Cas9-mediated deletion of full-length rat dystrophin. A Organization of the rat Dmd that spans ~ 2.3 mbp of the X chromosome. The 5 exons targeted for removal – 22–26 (in gray) and the two flanking exons (21 & 27 – in green) are shown in more detail (WT). The two guide RNAs (5’ and 3’ gRNAs) aided in the Cas9-mediated deletion of 18,307 bps that include exons 22–26 (MDR). B The 5’ and 3’ untranslated regions (unfilled magenta boxes) flank the 79 exons (filled magenta boxes) that contribute coding sequence of full-length dystrophin protein. Indicated with a closer view of exons 21–27 is the number of nucleotides each exon contributes (in parentheses). The 5 deleted exons (in gray) together contribute 800 bp of protein coding sequence, whose removal leads to a frame shift and a premature termination codon within the region of the transcript coded by exon 27 (*, MDR). C Immunoblotting confirms loss of full-length dystrophin (427 kDa; black arrow) and presence of predicted 106 kDa N-terminal fragment (gray double arrow) in 4 mo MDR gastrocnemius and heart lysates, and D immunofluorescence confirms loss of dystrophin staining at the sarcolemma of muscle fibers and cardiomyocytes (scale bars = 25 μm). E-F Immunoblotting was also performed for dystrophin glycoprotein complex (DGC) members β-dystroglycan (β-DG), α-dystrobrevin (α-DB), and syntrophins ( n = 3). Data are displayed as mean ± SEM with individual values and were analyzed using Student’s T-tests (α = 0.05; * p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: The following primary antibodies were used for immunofluorescence in the present study: dystrophin (1:100; MANEX1011B, Clone 1C7; DSHB), β-dystroglycan (1:100; MANDAG2, Clone 7A11; DSHB), utrophin (1:50; VP-U579; Vector Laboratories), α-sarcoglycan (1:50; VP-A105; Vector Laboratories); syntrophins (1:2000; #11425; Abcam), perilipin 1 (1:500; #9349; Cell Signaling Technology), anti-Laminin β2 antibody (1:10,000; [ ]), embryonic MHC (1:100; F1.652; DSHB), type I myofibers (1:50; BA-D5; DSHB), Type IIa myofibers (1:50; SC-71; DSHB), type IIx myofibers (1:20; 6H1; DSHB), pan-muscle MHC(1:100; MF 20; DSHB), COMP (1:800; #NBP2-92658; Noveus Biologicals), α-SMA (1:1000; #7817; Abcam), SMOC2 (1:500; #AF5140; R&D Systems), and PDGFRα (1:500; #AF1062; R&D Systems).

Techniques: CRISPR, Sequencing, Western Blot, Immunofluorescence, Staining

Journal: mBio

Article Title: Loss of LafB activity reverses daptomycin resistance in E. faecium

doi: 10.1128/mbio.00715-25

Figure Lengend Snippet: TLC analysis of glycolipids in total membrane lipid extracts from E. faecium . Glycolipid profiles of E. faecium SM1, SM1 lafB T577C, SM1Δ lafB , and SM1 Δ lafB /pAT28:: lafB mutants were compared to WT E. faecium SM1. Purified lipid standards included Gal₂DAG [1,2-diacyl-3-O-(α-D-galactosyl(1→6)-β-D-galactosyl)-sn-glycerol] and Glc ₁ DAG [1,2-diacyl-3-O-(α-D-glucopyranosyl)-sn-glycerol] (Avanti Polar Lipids). Glycolipid species Glc ₁ DAG, Glc ₂ DAG, and Gal ₂ DAG were identified based on migration relative to standards. Additional bands representing unknown lipid species are also indicated. The absence of glycolipid bands in the Δ lafB mutant and their restoration in the complemented strain demonstrates the role of lafB in glycolipid biosynthesis. Data are representative of three independent experiments.

Article Snippet: Approximately 5 μL of each of the two standards (Gal 2 DAG: 1,2-diacyl-3- O -(α- d -galactosyl1-6)-β- d -galactosyl- sn -glycerol Avanti Polar Lipids 840524P, Glc 1 DAG: 1,2-diacyl-3-O-(α-D-glucopyranosyl)-sn-glycerol Avanti Polar Lipids 840522P) and ~15 μL of each sample was spotted on the baseline of a Silica gel 60 TLC plate.

Techniques: Membrane, Purification, Migration, Mutagenesis