prolong gold antifade reagent with dapi  (Cell Signaling Technology Inc)

Journal: Journal of Functional Biomaterials

Article Title: A Borophosphate Glass Doped with Cobalt Oxide Improves Skeletal Muscle Structure and Function in Myopathic Mice

doi: 10.3390/jfb17030155

Figure Lengend Snippet: CoO-TRIM increased myofibers with centrally nucleated myofibers (CLN) and modulated proteolytic activity. ( A ) Representative images of TA muscle cross-sections at 14 dpt. Laminin (white): basal laminae; DAPI (blue): nuclei. Yellow arrows depicting position of central nuclei. ( B ) Summary values ( n = 4–5/group) for total myofibers with CLN normalized to TA muscle cross-section area (mm 2 ) at 14 dpt. ( C ) Summary values ( n = 4–5/group) for total myofibers (CLN + + CLN − ) normalized to TA muscle cross-section (mm 2 ) at 14 dpt. ( D ) Representative immunoblot for αII-Spectrin. The 145 kDa cleavage byproduct was normalized to total protein per lane, represented by the 40 kDa band from the total protein stain, and analyzed as a ratio of the 250 kDa band of αII-Spectrin. Mean densitometric data revealed a significant reduction in cleaved αII-Spectrin abundance in TA muscles of D2. mdx TRIM mice at 14 dpt. ( E ) Representative immunoblot of LC3B II and mean densitometric data at 14 dpt revealed reduced autophagosome number following CoO-TRIM administration in D2. mdx mice. ( n = 5/group); Summary values are means ± SEM. Comparisons made vs. WT and vehicle controls by 2-Way ANOVA, p < 0.05 = significant. Scale bars = 100 µm.

Article Snippet: Sections were washed 3× in TBS, incubated with secondary antibodies in blocking buffer for 60 min at RT, washed 3× in TBS, and mounted in InvitrogenTM ProLongTM Gold antifade reagent with DAPI (Cat.# P36941 , Fisher Scientific, Hampton, NJ, USA).

Techniques: Activity Assay, Western Blot, Staining, Muscles

Journal: Journal of Functional Biomaterials

Article Title: A Borophosphate Glass Doped with Cobalt Oxide Improves Skeletal Muscle Structure and Function in Myopathic Mice

doi: 10.3390/jfb17030155

Figure Lengend Snippet: Comparison of CLN in myofibers of WT and D2. mdx treated with Saline and TRIM mice. ( A , E ) Representative images of TA muscle cross-sections at 70 dpt and 140 dpt. Laminin (white): basal laminae; DAPI (blue): nuclei. Yellow arrows identify position of central nuclei. ( B , F ) Summary values for total myofibers with CLN (Top) and total myofibers (CLN + + CLN − ) (Bottom) normalized to TA muscle cross-section area (mm 2 ). Summary values presented for ( B ) 70 dpt and ( F ) 140 dpt. ( n = 7–8/group); scale bars = 100 µm. Proteolytic activity is altered following CoO-TRIM treatment. ( C , G ) Representative immunoblots for αII-Spectrin. The 145 kDa cleavage byproduct was normalized to total protein per lane, represented by the 40 kDa band from the total protein stain, and analyzed as a ratio of the 250 kDa band of αII-Spectrin. Mean densitometric data revealed a significant reduction in cleaved αII-Spectrin abundance in TRIM mice at 70 dpt. There was no difference at 140 dpt. ( D , H ) Representative immunoblots of LC3B II and mean densitometric data at ( D ) 70 dpt, revealed that TRIM reduced autophagosome number. No differences were observed at ( H ) 140 dpt. ( n = 7–8/group); summary values are means ± SEM. Comparisons made vs. vehicle controls by two-tailed Student’s t -test; p < 0.05 = significant.

Article Snippet: Sections were washed 3× in TBS, incubated with secondary antibodies in blocking buffer for 60 min at RT, washed 3× in TBS, and mounted in InvitrogenTM ProLongTM Gold antifade reagent with DAPI (Cat.# P36941 , Fisher Scientific, Hampton, NJ, USA).

Techniques: Comparison, Saline, Activity Assay, Western Blot, Staining, Two Tailed Test

Journal: Journal of Inflammation Research

Article Title: Integrated Multi-Omics Analysis Reveals IRF1-Driven Microglial PANoptosis via ZBP1 in Spinal Cord Injury

doi: 10.2147/JIR.S574990

Figure Lengend Snippet: IRF1 drove Zbp1 transcription in microglia. ( A ) Heatmap showing TF activity scores (right) and fold-change expression (left) associated with PANoptosis activity in microglia. ( B ) UMAP visualization of transcriptional activity scores (top) and gene expression levels (bottom) of Nr1d1, Spi1, Myc, Irf1 , and Cebpd across all cell types. ( C ) Heatmap showing the results of Pearson’s correlation analysis between Nr1d1, Spi1, Myc, Irf1 , and Cebpd and PANoptosis activity scores. ( D ) Bar graph showing the inferred TF activity at bulk RNA-seq levels. ( E ) The IGV browser illustrating the potential regulatory relationship between NR1D1, SPI1, MYC, IRF1, and CEBPD and the Zbp1 promoter region. ( F ) UCSC Genome Browser showing a distinct IRF1 binding peak within the Zbp1 promoter region, based on public ChIP-seq data. ( G ) Immunofluorescence at 7 days after SCI showing nuclear translocation of IRF1 (red) in Iba1+ microglia (green); nuclei were counterstained with DAPI (blue). Scale bars, 20μm (n=4 rats per group). ( H ) WB analysis of IRF1 and ZBP1 protein expression of microglia in vitro. Vinculin was used as a loading control. *, p<0.05; ****, p<0.0001.

Article Snippet: Nuclei were counterstained with DAPI (HY-K1048, MedChemExpress).

Techniques: Activity Assay, Expressing, Gene Expression, RNA Sequencing, Binding Assay, ChIP-sequencing, Immunofluorescence, Translocation Assay, In Vitro, Control