neural stem cells nscs  (Dawley Inc)

Journal: Frontiers in Pharmacology

Article Title: Astragaloside IV attenuates oxidative stress-associated mitophagy-dependent ferroptosis in peritoneal mesothelial cells via suppression of STAT3 signaling

doi: 10.3389/fphar.2026.1747615

Figure Lengend Snippet: AS-IV inhibits mitophagy to attenuate ferroptosis in PMCs by inhibiting STAT3 phosphorylation. For in vivo experiments (A–C) mice received daily intraperitoneal injection for 4 weeks with saline (Control), 4.25% dextrose PDF (100 mL/kg, PD), PD + AS-IV (15 mg/kg), or PD + S3I-201 (10 mg/kg). Tissues were collected at the end of week 4. For in vitro experiments, HMrSV5 cells were treated with 3% dextrose PDF for 24 h in the absence or presence of AS-IV (200 μM), colivelin (1 μM), or S3I-201 (100 μM). Groups for (D,E) Control, PD, PD + colivelin, PD + colivelin + AS-IV. Groups for panels (F,G) : Control, PD, PD + AS-IV, PD + S3I-201. (A) Representative HE staining, Masson staining, and immunohistochemical staining of peritoneal tissues. Scale bar, 60 μm. (B,C) Expression of p-STAT3, STAT3, PTGS2, PINK1, BNIP3, GPX4, and FUNDC1 in mouse peritoneal tissues was analyzed by Western blotting. Band intensities of PTGS2, PINK1, BNIP3, GPX4, and FUNDC1 were normalized to β-actin, and p-STAT3 was quantified as the p-STAT3/STAT3 ratio (n = 4). (D,E) Expression of p-STAT3, STAT3, PTGS2, PINK1, p62, BNIP3, GPX4, FUNDC1, and LC3B in HMrSV5 cells (Control, PD, PD + colivelin, PD + colivelin + AS-IV) was analyzed by Western blotting, with quantification as described above (n = 3). (F,G) Expression of p-STAT3, STAT3, PTGS2, PINK1, p62, BNIP3, GPX4, FUNDC1, and LC3B in HMrSV5 cells (Control, PD, PD + AS-IV, PD + S3I-201) was analyzed by Western blotting, with quantification as described above (n = 3). Data are presented as mean ± SD. One-way ANOVA with Tukey’s multiple-comparisons test was used. For (C , G) * p < 0.05, ** p < 0.01, *** p < 0.001 vs. PD. For (E) * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Control; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. PD + colivelin.

Article Snippet: The chemicals methylglyoxal (MGO; HY-w020014), S3I-201 (HY-15146), colivlin (HY-P1061), liproxstatin-1 (HY-12726), Carbonyl cyanide m-chlorophenylhydrazone (CCCP; HY-100941), erastin (HY-15763), N-acetylcysteine (NAC; HY-B0215), and mitochondrial division inhibitor 1 (Mdivi-1; HY-15886) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Phospho-proteomics, In Vivo, Injection, Saline, Control, In Vitro, Staining, Immunohistochemical staining, Expressing, Western Blot

Journal: Oncogene

Article Title: Early induction of the Rho-GEF ECT2 drives MEK/ERK oncogenic signaling in pancreatic ductal adenocarcinoma

doi: 10.1038/s41388-026-03860-3

Figure Lengend Snippet: A Dose–response curves showing cell viability of AsPC-1, CFPAC-1, MiaPaCa-2, and PANC-1 cells expressing shNT or shECT2 following treatment with increasing concentrations of Trametinib; n = 5. B Trametinib IC50 values in PDAC cell lines expressing shNT or shECT2. C Soft agar colony formation of PANC-1 cells expressing shNT or shECT2 treated with increasing concentrations of Trametinib; n = 5 * p < 0.05 relative to vehicle treated control. D Schematic representation of PKCι inhibition by ANF within the PKCι–ECT2–Rac1–PAK–MEK/ERK signaling pathway and immunoblot analysis of AsPC-1, CFPAC-1, MiaPaCa-2, and PANC-1 cells treated with ANF showing p-ECT2, ECT2, p-ERK1/2, and ERK1/2 expression. Dose–response curves showing cell viability of PDAC cell lines ( E ) and quantification of relative colony formation ( F ) following treatment with increasing concentrations of ANF; n = 5; * p < 0.05 relative to vehicle treated control. G Dose–response analysis of ANF and Trametinib alone or in combination in AsPC-1 cells; n = 5. H Combination index (CI) analysis of ANF and Trametinib in PDAC cells. CI < 0.9 indicates synergism, CI between 0.90 and 1.20 indicates additivity, and CI > 1.2 indicates antagonism. I Representative images and quantitation of colony formation of PANC-1 cells treated with ANF alone, Trametinib alone, or the combination of ANF and Trametinib; n = 5; * p < 0.05 relative to vehicle treated control. * * p < 0.05 relative to Trametinib alone treatment. J Immunoblot analysis of PDAC cell lines treated with the Rac1 inhibitor NSC23766 showing total and phosphorylated ERK1/2 status. Dose–response curves showing cell viability of PDAC cell lines ( K ) and quantification of relative colony formation ( L ) following treatment with increasing concentrations of with NSC23766; n = 5; * p < 0.05 relative to vehicle treated control. M Relative dose–response curves showing the combined effects of NSC23766 and Trametinib treatment in AsPC-1 cells; n = 5. N CI analysis showing the interaction between NSC23766 and Trametinib treatment on the viability of PDAC cells. O Representative images and quantitation of colony formation of PANC-1 cells treated with NSC23766 alone, Trametinib alone, or the combination of NSC23766 and Trametinib. * p < 0.05 relative to vehicle treated control. ** p < 0.05 relative to Trametinib alone treatment. For immunoblots, B-actin served as loading control and phospho-proteins were normalized to the corresponding total protein and expressed relative to vehicle treated cells.

Article Snippet: Trametinib, NSC23766 and Y-27632 were obtained from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Expressing, Control, Inhibition, Western Blot, Quantitation Assay

Journal: Bioactive Materials

Article Title: Development of a PTEN -siRNA activated scaffold to promote axonal regrowth following spinal cord injury

doi: 10.1016/j.bioactmat.2026.01.022

Figure Lengend Snippet: PTEN -siRNA nanoparticle transfection enables efficient gene silencing with minimal impact on neuron health (A) PTEN -siRNA nanoparticles (100 nM) demonstrated highly efficient knockdown of target PTEN mRNA in differentiated NSC34 neurons, with peak knockdown on Day 3 for the delivered siRNA [t(2) = −4.840, p = 0.02007; one-tailed] with expression levels returning to baseline by Day 7 [t(2) = 0.05373, p = 0.5190; one-tailed]. RNA fold change and associated error was assessed using the 2 −ΔΔCt method and normalized to GAPDH as per (B) Treatment of neurons with PTEN- siRNA nanoparticles at 100 nM had a non-significant effect on the metabolic activity at 3 [t(2) = 0.6370, p = 0.5893, 95 % CI 74.4820 0.1; • = p ≤ 0.1, ∗ = p ≤ 0.05, ∗∗ = p ≤ 0.01, ∗∗∗ = p ≤ 0.001, ∗∗∗∗ = p ≤ 0.0001.

Article Snippet: NSC34 neuronal culture and transfections: NSC34 neurons (Cedarlane Cat No. CLU140), a hybrid cell line produced by the fusion of motor neuron enriched embryonic mouse spinal cord cells with mouse neuroblastoma cells, were cultured in growth medium Dulbecco's Modified Eagle Medium (DMEM; Sigma-Aldrich, Ireland) supplemented with 10 % fetal bovine serum (FBS; Labtech UK), 1 % (v/v) L-Glutamine (Sigma-Aldrich, Ireland), and 1 % (v/v) Penicillin- Streptomycin Solution (Sigma-Aldrich, Ireland) in a T-175 cell culture flask (37 °C, 5 % CO 2 ).

Techniques: Transfection, Knockdown, One-tailed Test, Expressing, Activity Assay, Control, Staining

Journal: Bioactive Materials

Article Title: Development of a PTEN -siRNA activated scaffold to promote axonal regrowth following spinal cord injury

doi: 10.1016/j.bioactmat.2026.01.022

Figure Lengend Snippet: Scaffold-mediated siRNA delivery does not affect the viability of neurons, results in prolonged downregulation of PTEN, and stimulates increased expression of BCL2 and GAP43 mRNA . (A) The metabolic activity of NSC34 neurons cultured on PTEN -siRNA activated scaffolds peaked by day 3 post-transfection [t(2) = 4.0599, p = 0.05565, 95 % CI 93.954 0.1; • = p ≤ 0.1, ∗ = p ≤ 0.05, ∗∗ = p ≤ 0.01, ∗∗∗ = p ≤ 0.001, ∗∗∗∗ = p ≤ 0.0001.

Article Snippet: NSC34 neuronal culture and transfections: NSC34 neurons (Cedarlane Cat No. CLU140), a hybrid cell line produced by the fusion of motor neuron enriched embryonic mouse spinal cord cells with mouse neuroblastoma cells, were cultured in growth medium Dulbecco's Modified Eagle Medium (DMEM; Sigma-Aldrich, Ireland) supplemented with 10 % fetal bovine serum (FBS; Labtech UK), 1 % (v/v) L-Glutamine (Sigma-Aldrich, Ireland), and 1 % (v/v) Penicillin- Streptomycin Solution (Sigma-Aldrich, Ireland) in a T-175 cell culture flask (37 °C, 5 % CO 2 ).

Techniques: Expressing, Activity Assay, Cell Culture, Transfection, Control, Lactate Dehydrogenase Assay, Gene Expression, One-tailed Test