phospho mk2  (Cell Signaling Technology Inc)

Journal: Journal of Translational Medicine

Article Title: From injury to recovery: transcriptomic dynamics in zebrafish brain regeneration

doi: 10.1186/s12967-025-07400-7

Figure Lengend Snippet: Transcriptomic and pathway insights into zebrafish brain regeneration. A ) Gene Ontology (GO) enrichment analysis of biological processes (BP), molecular functions (MF), and cellular components (CC) across the three regenerative stages (1, 4, and 7 days post-lesion; dpl). The vertical axis lists the enriched GO terms, while the horizontal bars represent the gene ratios associated with each term. The depth of the color indicates the adjusted p-value, and the circle size corresponds to gene counts contributing to each term. B ) heatmap showing the expression levels of key cell cycle-related genes at 1, 4, and 7 dpl. Stage-specific activation patterns, such as peak expression of proliferation markers (aurka, aurkb, wdr76, etc.) at 4 dpl and sustained expression of MCM proteins, reflect dynamic cellular processes during brain regeneration. C ) overview of KEGG pathways enriched across the three regeneration stages. The visualization underscores the pathways driving key biological processes during zebrafish brain regeneration. D ) heatmaps showing the expression of genes related to the MAPK signaling pathway: i ) upregulated genes and ii ) downregulated genes, across the three stages, emphasizing the dynamic regulation of this critical pathway. E ) immunoblot analysis showing the levels of phosphorylated (active) and total p38α MAPK protein during zebrafish brain regeneration. F ) immunoblot analysis showing the levels of phosphorylated (active) and total MK2 and Hsp27 along with c-Jun protein during zebrafish brain regeneration. β-actin was used as the loading control for both E-F panels. G ) quantification of phosphorylated (active) and total p38α MAPK levels during regeneration is shown as a bar graph ( n = 3). H ) quantification of phosphorylated (active) and total MK2 levels during regeneration is shown as a bar graph ( n = 3). I ) quantification of c-Jun levels during regeneration is shown as a bar graph ( n = 3). J ) quantification of phosphorylated (active) and total HSP27 levels during regeneration is shown as a bar graph ( n = 3). Significance is represented as n.s. for p-value > 0.05, * for p-value < 0.05, ** for p-value < 0.01 and *** for p-value < 0.001. dpl: days post-lesion

Article Snippet: Rabbit monoclonal Phospho p38 MAPK (1:1000; 4511S; Cell Signaling Technology), rabbit polyclonal total p38 MAPK (1:1000; 9212S; Cell Signaling Technology), rabbit monoclonal Phospho HSP27 (1:1000; 9709S; Cell Signaling Technology), mouse monoclonal Total HSP27 (1:500; CPTC-HSPB1-2; Developmental Studies Hybridoma Bank), rabbit monoclonal Phospho MK2 (1:1000; 3007S; Cell Signaling Technology), Total MK2 (1:1000; EB11929; Everest Biotech), rabbit monoclonal c-Jun (1:1000; 9165S; Cell Signaling Technology), mouse monoclonal β-catenin (1:1000; 610153; BD Biosciences), Phospho AKT (1:1000; 4058S; Cell Signaling Technology), Total AKT (1:1000; 2920S; Cell Signaling Technology), mouse monoclonal β-actin (1:1000; sc47778; Santa Cruz).

Techniques: Expressing, Activation Assay, Western Blot, Control

Journal: Journal of Translational Medicine

Article Title: From injury to recovery: transcriptomic dynamics in zebrafish brain regeneration

doi: 10.1186/s12967-025-07400-7

Figure Lengend Snippet: BIRB796-mediated inhibition of MAPK14a pathway impairs the regenerative response post traumatic brain injury. A ) schematic showing BIRB796 treatment after zebrafish brain injury and the brains harvested at 4 dpl. Treatment regimen: animals were immersed in system water containing 0.5 μM or 1 μM BIRB796 (or vehicle control: 0.1% DMSO) immediately after lesion induction and maintained until sacrifice at 4 dpl. B ) immunoblot analysis showing the levels of phosphorylated (active) and total p38α MAPK protein upon BIRB796 treatment at 4 dpl;. *represents the band for p38α MAPK. C ) quantification of phosphorylated (active) and total p38α MAPK levels in control regenerating (4 dpl DMSO) brain and BIRB796 treated regenerating (4 dpl) brain is shown as a bar graph ( n = 3). D ) immunoblot analysis showing the levels of phosphorylated (active) and total Hsp27 protein upon BIRB796 treatment at 4 dpl E ) quantification of phosphorylated (active) and total Hsp27 levels in control regenerating (4 dpl DMSO) brain and BIRB796 treated regenerating (4 dpl) brain is shown as a bar graph ( n = 3). F ) immunoblot analysis showing the levels of phosphorylated (active) and total MK2 and c-Jun protein upon BIRB796 treatment at 4 dpl. G ) quantification of phosphorylated (active)and total MK2 levels in control regenerating (4 dpl DMSO) brain and BIRB796 treated regenerating (4 dpl) brain is shown as a bar graph ( n = 3). H ) quantification of c-Jun levels in control regenerating (4 dpl DMSO) brain and BIRB796 treated regenerating (4 dpl) brain is shown as a bar graph ( n = 3). Significance is represented as n.s. for p-value > 0.05, * for p-value < 0.05, ** for p-value < 0.01 and *** for p-value < 0.001. ( I ) Immunofluorescence staining on 4 dpl zebrafish brains treated with DMSO (control) or 1 μM BIRB796. Panels show staining with the following markers: PCNA (red), HuC/D (red), GFAP (green), and S100B (green), with DAPI used to label nuclei. ( A - D ) DMSO control, (A’-D’) BIRB796 treatment. Scale bar denotes 10 μm. The following anatomical regions are indicated: Dc (central zone of the dorsal telencephalic area), Dm (medial zone of dorsal telencephalic area), Dl (lateral zone of dorsal telencephalic area), Vd (dorsal nucleus of ventral telencephalic area). ( J - M ) quantification of immunostaining wherein injured hemisphere of 4 dpl regenerating brain has been compared to that of 4 dpl BIRB796 treated regenerating brain ( n = 3). Significance is represented as n.s. for p-value > 0.05, * for p-value < 0.05, ** for p-value < 0.01 and *** for p-value < 0.001. cntrl: control; dpl: days post-lesion

Article Snippet: Rabbit monoclonal Phospho p38 MAPK (1:1000; 4511S; Cell Signaling Technology), rabbit polyclonal total p38 MAPK (1:1000; 9212S; Cell Signaling Technology), rabbit monoclonal Phospho HSP27 (1:1000; 9709S; Cell Signaling Technology), mouse monoclonal Total HSP27 (1:500; CPTC-HSPB1-2; Developmental Studies Hybridoma Bank), rabbit monoclonal Phospho MK2 (1:1000; 3007S; Cell Signaling Technology), Total MK2 (1:1000; EB11929; Everest Biotech), rabbit monoclonal c-Jun (1:1000; 9165S; Cell Signaling Technology), mouse monoclonal β-catenin (1:1000; 610153; BD Biosciences), Phospho AKT (1:1000; 4058S; Cell Signaling Technology), Total AKT (1:1000; 2920S; Cell Signaling Technology), mouse monoclonal β-actin (1:1000; sc47778; Santa Cruz).

Techniques: Inhibition, Control, Western Blot, Immunofluorescence, Staining, Immunostaining

Journal: Viruses

Article Title: The Combination of GS-441524 (Remdesivir) and Ribavirin Results in a Potent Antiviral Effect Against Human Parainfluenza Virus 3 Infection in Human Airway Epithelial Cell Cultures and in a Mouse Infection Model.

doi: 10.3390/v17020172

Figure Lengend Snippet: Figure 1. The combined antiviral effect of ribavirin with either remdesivir or GS-441524 results in synergistic antiviral efficacy against HPIV-3 replication in LLC-MK2 cells. (A,D) The matrices of inhibition of HPIV-3 replication in the presence of combined treatments. Data are from five independent experiments. (B,E) The interaction between two molecules was determined by using the Bliss independence model. Data are presented as mean values of BLISS scores. Data are analyzed using the one-sample Student’s t-test. p < 0.05, *; p < 0.01, **; p < 0.001, ***. (C,F) 3D synergy maps.

Article Snippet: Ribavirin used in LLC-MK2 cell and human airway epithelial cell cultures was purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Inhibition