Journal: Journal of Molecular and Cellular Cardiology Plus

Article Title: Independent and synergistic roles of MEK-ERK1/2 and PKC pathways in regulating functional changes in vascular tissue following flow cessation

doi: 10.1016/j.jmccpl.2025.100300

Figure Lengend Snippet: Signaling pathways involved in ET B receptor upregulation and VSMC contraction. Extracellular stimuli such as flow cessation activate the MAPK/ERK pathway, which is disrupted by MEK inhibition (Trametinib) or ERK inhibition (Ulixertinib). In parallel, intracellular Ca 2+ increase leads to delayed PKC activation, promoting NF-κB signaling via the IKK complex. PKC and NF-κB inhibition (RO-317549 and BMS345541, respectively) remain effective when applied up to 6 h post-stimulus. Combined inhibition further suppresses ET B upregulation on VSMCs and reduces receptor-mediated contraction, indicating that both pathways contribute independently and are required for full functional response . Abbreviations: ET B : endothelin type B receptor; VSMC: vascular smooth muscle cell; NF-κB: nuclear factor. Created in BioRender. Kazantzi, S. (2025) https://BioRender.com/u89c445

Article Snippet: The membranes were then transferred to a rocking table in room temperature with Blocking Buffer (EveryBlot Blocking Buffer, Cat. #: 12010020, Bio-Rad) for 10 min and then, the membranes were incubated with primary antibody dilutions overnight in 4 °C (1:1000 Mouse Anti-p44 / 42 MAP Kinase Monoclonal Antibody, Unconjugated, Clone L34F12 [Cell Signaling Technology Cat# 4696, lot: 29, RRID: AB_390780 ], 1:1000 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb [Cell Signaling Technology Cat# 4370, Lot: 28, RRID: AB_2315112 ), 1:1000 NF-κB p65 (L8F6) Mouse mAb [Cell Signaling Technology Cat# 6956, Lot:10, RRID: AB_10828935 ], 1:1000 Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb [Cell Signaling Technology Cat# 3033,Lot: 19, RRID: AB_331284 ], GAPDH (D16H11) XP Rabbit mAb, [Cell Signaling Technology Cat# 5174, Lot: 8, RRID: AB_10622025 ], 1:1000 Actin, Smooth Muscle (1A4) Mouse Monoclonal Antibody [Cell Marque Cat# 202 M-94, Lot:0000243213, RRID: AB_1157937 ], 1:1000 alpha smooth muscle Actin antibody (EPR5368) [Abcam Cat# ab124964, Lot: GR303485 –26, RRID: AB_11129103 ]).

Techniques: Protein-Protein interactions, Inhibition, Activation Assay, Functional Assay

Journal: Cells

Article Title: Small Extracellular Vesicles Promote Axon Outgrowth by Engaging the Wnt-Planar Cell Polarity Pathway

doi: 10.3390/cells14010056

Figure Lengend Snippet: List of antibodies.

Article Snippet: Mouse anti-MAP-2 (Clone AP20, 1:500) , Millipore (Darmstadt, Germany) , MAB3418.

Techniques: Transduction

Journal: Cell reports

Article Title: Cofilactin rod formation mediates inflammation-induced neurite degeneration

doi: 10.1016/j.celrep.2024.113914

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse anti-Microtubule associated protein (MAP-2) , , Millipore-Sigma , Cat# MAB3418, RRID: AB_11212326.

Techniques: Affinity Purification, Recombinant, Membrane, Software

Journal: Cell reports

Article Title: Cofilactin rod formation mediates inflammation-induced neurite degeneration

doi: 10.1016/j.celrep.2024.113914

Figure Lengend Snippet: CAR formation in neuron-glia co-cultures is attenuated by both neuronal cofilin-1 hemizygosity and glial p47 phox deficiency (A) Representative photomicrographs of co-cultures immunostained for microtubule-associated protein-2 (MAP-2) (blue) and CARs (green). Scale bar: 10 μm. (B–D) CAR density expressed as percentage of total neurite area. n = 4; *p < 0.05 and **p < 0.01 vs. control by one-way ANOVA with Dunnett’s test. All data are shown as mean ± SEM.

Article Snippet: Primary antibodies were obtained from the following sources and used at 1:500 dilutions: rabbit anti-cofilin-1, Cytoskeleton #ACFL02, Denver, CO; mouse anti-MAP-2, Millipore #3418, Temecula, CA; mouse anti-NeuN, Millipore #MAB377B, Temecula, CA; mouse anti-NF-H, BioLegend #SMI31, San Diego, CA; rabbit anti-Phospho-Histone H2AX (Ser139) Antibody #2577; rabbit Iba1, Wako 019–19741.

Techniques:

Journal: Cell reports

Article Title: Cofilactin rod formation mediates inflammation-induced neurite degeneration

doi: 10.1016/j.celrep.2024.113914

Figure Lengend Snippet: Neurite loss in neuron-glia co-cultures is attenuated by neuronal cofilin-1 hemizygosity and glial p47 phox deficiency (A) Photomicrographs of co-cultures immunostained for MAP-2 (green) and NeuN (blue). Scale bar: 20 μm. (B–D) Neurite length assessed at 4, 24, and 48 h of 50 ng/mL S100β incubation, expressed relative to control wells of the respective co-culture type. n = 4; *p < 0.05 and **p < 0.01 by one-way ANOVA with Dunnett’s test. (E) Mechanism proposed for inflammation-induced neurite degeneration. In response to pro-inflammatory stimuli, brain microglia and infiltrating macrophages upregulate superoxide production by NADPH oxidase. Resulting oxidative stress in nearby neurites leads to formation of CARs. Persistence of the CARs causes neurite degeneration, which can occur in the absence of parental neuron death. This process is attenuated in p47 phox−/− mice, which cannot form an active NADPH oxidase-2 complex, and in cofilin hemizygous ( COF −/+ ) mice, which have reduced propensity to form CARs. All data are shown as mean ± SEM.

Article Snippet: Primary antibodies were obtained from the following sources and used at 1:500 dilutions: rabbit anti-cofilin-1, Cytoskeleton #ACFL02, Denver, CO; mouse anti-MAP-2, Millipore #3418, Temecula, CA; mouse anti-NeuN, Millipore #MAB377B, Temecula, CA; mouse anti-NF-H, BioLegend #SMI31, San Diego, CA; rabbit anti-Phospho-Histone H2AX (Ser139) Antibody #2577; rabbit Iba1, Wako 019–19741.

Techniques: Incubation, Co-Culture Assay