311 in cyp11b2  (Vector Laboratories)

Journal: Cell Communication and Signaling : CCS

Article Title: Intercellular communication from myotubes to fibroblasts is differentially impacted by mineralocorticoid receptor signaling and muscular dystrophy

doi: 10.1186/s12964-025-02515-2

Figure Lengend Snippet: CM from myotubes induced gene expression changes in neonatal skeletal muscle fibroblasts. A Schematic of the experiment performed for bulk RNA sequencing of myotubes and fibroblasts. Primary myogenic progenitor cells and fibroblasts isolated from neonatal (< 5-day-old) C57 (wild-type) and mdx (DMD model) mice were used. Myotubes differentiated for 10 days were treated for 48 h with either Aldo (1 µM) or vehicle (Veh, DMSO), followed by 15 h of incubation in fresh media. This media (CM) was then collected and used to treat the muscle fibroblasts isolated from the same mouse genotype for 24 h. Myotubes and fibroblasts were then harvested and used for RNA sequencing. B Stacked bar plots demonstrating differential expression (DE) of genes comparing Aldo to Veh treatment and C57 (wild-type) to mdx (disease) in myotubes and CM-treated fibroblasts. For DE, DESeq2 in NovoMagic was used with a threshold of padj < 0.1 and |fold change|≥ 1.5. n = 3 biological replicates (mice) used per group for fibroblast and myoblast isolations; 4 technical replicates per mouse (fibroblasts) combined for RNA isolation ( C ) Representative images (left) and quantification (right) of immunohistochemistry of aldosterone synthase (Cyp11b2) in 2-day-old C57 and mdx hind-limb muscle, showed increased Cyp11b2 in mdx muscle. T-tests were used for statistical analysis, and significance was established at p < 0.05. Scale bar = 100 µm. * indicates statistical significance

Article Snippet: Briefly, sections were incubated in Cyp11b2 (1:250, AS2084, rabbit anti-mouse) overnight at 4 °C, and then in HRP-conjugated goat anti-rabbit (1:200, 111–035–144, Jackson ImmunoResearch Labs) at room temperature for 1 h, followed by detection with ImmPACT DAB (SK-4105, Vector).

Techniques: Gene Expression, RNA Sequencing, Isolation, Incubation, Quantitative Proteomics, Immunohistochemistry

Journal: Antioxidants

Article Title: Identification of a Novel Inhibitor of TfR1 from Designed and Synthesized Muriceidine A Derivatives

doi: 10.3390/antiox11050834

Figure Lengend Snippet: 12 b bind to a glycoprotein containing α, β-D galactose. ( A , B ) Effect of 12b on RTK proteins stabilization. MDA-MB-231 cells were treated with or without 12b (4 μM) for 2 h before being heated at different temperatures. Protein levels were analyzed by Western blotting. ( C , D ) The relative band intensity to β-Tubulin was analyzed by ImageJ software. Protein band densities were quantified by normalizing to DMSO. ( E ) The samples in ( A ) were taken for immunoblotting, membranes were incubated with the RCA I antibody, and HRP-Streptavidin antibody. Signals were detected by chemiluminescence.

Article Snippet: The membranes were incubated with the indicated RCA I antibody (Vector Laboratories, Inc., Burlingame, CA, USA) for 30 min at room temperature, followed by HRP-Streptavidin antibody (R&D Systems, Minneapolis, MN, USA) at room temperature for 1 h. After washing with TBST, signals were detected by chemiluminescence with the enhanced chemiluminescence (ECL) detection reagents.

Techniques: Western Blot, Software, Incubation

Journal: Histochemistry and Cell Biology

Article Title: Astroglial and oligodendroglial markers in the cuprizone animal model for de- and remyelination

doi: 10.1007/s00418-022-02096-y

Figure Lengend Snippet: Immunohistochemical analysis of S100B and RCA-1 (marker for activated microglia) expression. Representative pictures demonstrate double immunofluorescence staining with S100B (red) and ricinus communis agglutinin-1 (RCA-1) (green) performed in the central corpus callosum ( a ), cortex (represented layers V–VI in ( b ), and hippocampal areas CA3 ( c ) and dentate gyrus ( d ) of C57BL/6 mice at week 5 under cuprizone diet (severe demyelination). Co-localization of S100B with RCA-1 was not found. Scale bars: 10 µm ( a , d ), 20 µm ( b , c )

Article Snippet: Slides were then incubated overnight at 4 °C with the following primary antibodies: myelin proteolipid protein (PLP; mouse IgG2a, 1:500, Serotec, Bio-Rad), glial fibrillary acidic protein (GFAP; mouse IgG1, 1:200, Merck Millipore), (GFAP; rabbit, 1:200, Dako), vimentin (VIM, rabbit IgG,1:500, Abcam), aldehyde dehydrogenase (ALDH1L1, mouse IgG, 1:1000, Merck Millipore), calcium-binding protein B (S100B; rabbit IgG, 1:500, Abcam), adenomatous polyposis coli (APC, clone CC1; mouse IgG2b, 1:200, Calbiochem, Merck Millipore), lipocalin 2 (LCN2; goat IgG, 1:200, R&D), ricinus communis agglutinin (RCA-1, 1:1000, Vector Laboratories), OLIG2 (rabbit IgG,1:500, Chemicon, Merck Millipore) (Table ).

Techniques: Immunohistochemical staining, Marker, Expressing, Double Immunofluorescence Staining