Journal: Biochemistry and Biophysics Reports

Article Title: Co-loaded simvastatin-ginsenoside Rh2 liposomes enhance cellular immune responses as vaccine adjuvants

doi: 10.1016/j.bbrep.2025.102159

Figure Lengend Snippet: The characterizations of simvastatin(SIM) and ginsenoside Rh2-loaded liposome (LIPO-SIM-Rh2). (A) LIPO-SIM-Rh2 in PBS. (B) LIPO-SIM-Rh2 prepared by ethanol injection combined with rotary evaporation. (C) LIPO-SIM prepared by ethanol injection combined with rotary evaporation. (D) LIPO-SIM-Rh2 prepared by ethanol injection combined with rotary evaporation. (E) ζ-average size. (F) TEM micrograph of LIPO-SIM-Rh2. (G) Zeta potential. (H) Polydispersity index (PDI). (I) DLS (dynamic light scattering) (volume distribution). The data were presented as the mean ± standard deviation (SD) of results from three independent experiments.

Article Snippet: Simvastatin was procured from MedChemExpress (New Jersey, USA), while 20(S)-Ginsenoside Rh2 (with a purity of ≥98 %) was obtained from Macklin (Shanghai, China).

Techniques: Injection, Evaporation, Zeta Potential Analyzer, Standard Deviation

Journal: Cell death & disease

Article Title: Statins suppress cell-to-cell propagation of α-synuclein by lowering cholesterol.

doi: 10.1038/s41419-023-05977-9

Figure Lengend Snippet: Fig. 1 High-content drug screening for chemical modifiers of α-synuclein and mHtt propagation. A Schematic workflow of drug screening in BiFC cell models. B Heat map representing the mean percentage changes in aggregate propagation. Details are presented in Supplementary Table 1. C, D Comparison of secondary screening results for α-synuclein and mHtt. Compounds with shared inhibitory effects on α-synuclein and mHtt propagation are presented as a Venn diagram (C) and heat map (D). Details are presented in Supplementary Tables 2 and 3. E Concentration-response relationship for pravastatin (PRV). F Summary data showing that both PRV and simvastatin (SIMV) significantly reduced α-synuclein propagation in a concentration-dependent manner. Data are presented as means ± SEM (*p < 0.05, #p < 0.05, **p < 0.005, ##p < 0.005, ***p < 0.0005, ###p < 0.0005, ****p < 0.0001, ####p < 0.0001; one-way ANOVA with Dunnett’s post-hoc test).

Article Snippet: Briefly, adult (6-month old) transgenic mice and littermate WT mice were administered statins (pravastatin [Tocris 2318], simvastatin [Tocris 1965]; 1 mg/kg/d) daily for 3 months by oral gavage.

Techniques: Drug discovery, Comparison, Concentration Assay

Journal: Cell death & disease

Article Title: Statins suppress cell-to-cell propagation of α-synuclein by lowering cholesterol.

doi: 10.1038/s41419-023-05977-9

Figure Lengend Snippet: Fig. 2 Simvastatin significantly alleviates motor deficits and synucleinopathy lesions, whereas pravastatin does not. A, C Forelimb grip strength test. B, D Balance beam test. A, B Pravastatin administration. C, D Simvastatin administration. E, F Immunohistochemical analysis of pS129 in the motor cortex (M.ctx) and parietal cortex (pacx) of A53T mice treated with pravastatin (E) or simvastatin (F). Scale bar: 50 μm. A–F n = 11–15 mice per group. All data are presented as means ± SEM (*p < 0.05, **p < 0.005, ***p < 0.0005, ****p < 0.0001; one-way ANOVA with Dunnett’s post-hoc test).

Article Snippet: Briefly, adult (6-month old) transgenic mice and littermate WT mice were administered statins (pravastatin [Tocris 2318], simvastatin [Tocris 1965]; 1 mg/kg/d) daily for 3 months by oral gavage.

Techniques: Immunohistochemical staining

Journal: Cell death & disease

Article Title: Statins suppress cell-to-cell propagation of α-synuclein by lowering cholesterol.

doi: 10.1038/s41419-023-05977-9

Figure Lengend Snippet: Fig. 5 Cholesterol levels regulate α-synuclein aggregation and secretion. A, B Accumulation of α-synuclein aggregates (A) and secretion α- synuclein (B) in NPC1-KO and WT cells. C Total intracellular cholesterol levels in WT cells treated with or without U18. D, E Secretion (D) and intracellular accumulation (E) of α-synuclein aggregates in WT cells treated with and without U18. F–H Total cholesterol levels (F), intracellular accumulation of α-synuclein aggregates (H), and secretion of α-synuclein aggregates (G) in NPC1-KO cells treated with or without MBCD. I–K Intracellular total cholesterol (I), α-synuclein secretion (J), and intracellular α-synuclein aggregates (K) in NPC1-KO cells treated with or without pravastatin. L–N Intracellular total cholesterol (L), α-synuclein secretion (M), and intracellular α-synuclein aggregates (N) in NPC1-KO cells treated with or without simvastatin. All cell experiments were repeated triplicate determination of each sample. All data are presented as means ± SEM (*p < 0.05, **p < 0.005, ***p < 0.0005, ****p < 0.0001; two-tailed unpaired Student’s t-test [A–H], one-way ANOVA with Dunnett’s post-hoc test [I–N]).

Article Snippet: Briefly, adult (6-month old) transgenic mice and littermate WT mice were administered statins (pravastatin [Tocris 2318], simvastatin [Tocris 1965]; 1 mg/kg/d) daily for 3 months by oral gavage.

Techniques: Two Tailed Test

Journal: Cell death & disease

Article Title: Statins suppress cell-to-cell propagation of α-synuclein by lowering cholesterol.

doi: 10.1038/s41419-023-05977-9

Figure Lengend Snippet: Fig. 7 Effects of HFD feeding and simvastatin on motor functions and synucleinopathy lesions. A Experimental timeline of simvastatin administration and behavioral tests. B Forelimb grip strength test. C Balance beam test. D Immunohistochemical analyses of pS129 in the striatum, motor cortex (M.ctx), and rhinal cortex (Rh.ctx). E HFD-induced cholesterol accumulation in different brain regions, with and without simvastatin treatment. Scale bar: 20 μm. All data are presented as means ± SEM (*p < 0.05, **p < 0.005, ***p < 0.0005, ****p < 0.0001; two-way ANOVA with Tukey’s post-hoc test [B, C], one-way ANOVA with Dunnett’s post-hoc test [D, E]). All mice and brain tissues from mice were run individually with groups of 9 to 10 mice.

Article Snippet: Briefly, adult (6-month old) transgenic mice and littermate WT mice were administered statins (pravastatin [Tocris 2318], simvastatin [Tocris 1965]; 1 mg/kg/d) daily for 3 months by oral gavage.

Techniques: Immunohistochemical staining

Journal: Cell Death & Disease

Article Title: MiR-122-5p regulates the mevalonate pathway by targeting p53 in non-small cell lung cancer

doi: 10.1038/s41419-023-05761-9

Figure Lengend Snippet: A The proliferation of A549 cells treated with 122-5p inh. combined with si-p53 or 122-5p inh. alone or 122-5p mim. combined with Nutlin-3 or 122-5p mim. alone using the CCK-8 assay. The colony formation assay could detect the long-term proliferation ability of A549 cells treated with 122-5p inh. combined with si-p53 or 122-5p inh. alone or 122-5p mim. combined with Nutlin-3 or 122-5p mim. alone ( B ) and the bar graph corresponding to B ( D ). The migration ability of A549 cells treated with 122-5p inh. combined with si-p53 or 122-5p inh. alone or 122-5p mim. combined with Nutlin-3 or 122-5p mim. alone using the cell scratch assay ( C ) and the bar graph corresponding to C ( E ). The apoptosis of A549 cells treated with 122-5p inh. combined with si-p53 or 122-5p inh. alone or 122-5p mim. combined with Nutlin-3 or 122-5p mim. alone using flow cytometry ( F ) and the analysis of early apoptosis ( G ) and late apoptosis ( H ). The above bar graphs were the sum of three independent experiments (mean ± SD). The ctrl group was represented by A549 cells treated using a medium solution containing 122-5p inh. NC and 122-5p mim. NC and si-p53 NC and DMSO, ns: not significant, * P < 0.05 and ** P < 0.01 vs . Ctrl group, # P < 0.05 and ## P < 0.01 vs. 122-5p inh. group, & P < 0.05 and && P < 0.01 vs. 122-5p mim. group.

Article Snippet: Nutlin-3 (Glpbio, cat#GC16051) and Simvastatin (SIM, Selleck, cat#S1792) powder were dissolved in DMSO to prepare a stock solution.

Techniques: CCK-8 Assay, Colony Assay, Migration, Wound Healing Assay, Flow Cytometry

Journal: Cell Death & Disease

Article Title: MiR-122-5p regulates the mevalonate pathway by targeting p53 in non-small cell lung cancer

doi: 10.1038/s41419-023-05761-9

Figure Lengend Snippet: A549 cells treated with si-p53 or Nutlin-3, RT-qPCR could detect the relative mRNA expression of p53 ( A ) and ABCA1 ( B ). p53, ABCA1, SREBP2(P), SREBP2(M) protein expression using western blotting ( C ) and the bar graph corresponding to C after statistical analysis ( D ) in A549 cells treated with si-p53 or Nutlin-3. E The relative mRNA expression of the MVA pathway genes HMGCS1 ( F ), HMGCR ( G ), MVK ( H ), PMVK ( I ), MVD ( J ), FDPS ( K ), FDFT1 ( L ), SQLE ( M ), and GGPS1 ( N ) by RT-qPCR in A549 cells treated with Nutlin-3 combined with DL-MVA or Nutlin-3 alone or DL-MVA alone. The expression of the MVA pathway key proteins HMGCS1, HMGCR, and FDFT1 using western blotting ( O ) and the bar graph corresponding to O ( P ) in A549 cells treated with Nutlin-3 combined with DL-MVA or Nutlin-3 alone or DL-MVA alone. The above bar graphs were the sum of 3 independent experiments (mean ± SD). The ctrl group was A549 cells treated with a medium solution containing si-p53 NC and DMSO, ns: not significant, * P < 0.05 and ** P < 0.01 vs. Ctrl or DMSO group, # P < 0.05 and ## P < 0.01 vs . DL-MVA group.

Article Snippet: Nutlin-3 (Glpbio, cat#GC16051) and Simvastatin (SIM, Selleck, cat#S1792) powder were dissolved in DMSO to prepare a stock solution.

Techniques: Quantitative RT-PCR, Expressing, Western Blot

Journal: Cell Death & Disease

Article Title: MiR-122-5p regulates the mevalonate pathway by targeting p53 in non-small cell lung cancer

doi: 10.1038/s41419-023-05761-9

Figure Lengend Snippet: A The changes of A-CoA in A549 cells treated with Nutlin-3 combined with DL-MVA or Nutlin-3 alone by ELISA kit. Nutlin-3 combined with DL-MVA or Nutlin-3 alone changes mevalonic acid ( B ) and 25-Hydroxyergosterol ( D ) by LC-MS in treated A549 cells. C The changes of TC in A549 cells treated with Nutlin-3 combined with DL-MVA or Nutlin-3 alone by biochemical kit. ( E ) The changes of A-CoA in A549 cells treated with 122-5p inh. combined si-p53 or 122-5p inh. alone using an ELISA kit. The changes of mevalonic acid ( F ) and 25-hydroxyergosterol ( H ) in A549 cells. G The changes of TC in A549 cells treated with 122-5p inh. combined with si-p53 or 122-5p inh alone by biochemical kit. The ctrl group was A549 cells treated with a medium solution containing 122-5p inh. NC and si-p53 NC. The above bar graphs were the sum of three independent experiments (mean ± SD), ns: not significant, * P < 0.05 and ** P < 0.01 vs DMSO or Ctrl group, # P < 0.05 and ## P < 0.01 vs Nutlin-3 or 122-5p inh. group.

Article Snippet: Nutlin-3 (Glpbio, cat#GC16051) and Simvastatin (SIM, Selleck, cat#S1792) powder were dissolved in DMSO to prepare a stock solution.

Techniques: Enzyme-linked Immunosorbent Assay, Liquid Chromatography with Mass Spectroscopy

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Targeting Cholesterol Metabolism as Efficient Antiviral Strategy Against the Hepatitis E Virus

doi: 10.1016/j.jcmgh.2021.02.002

Figure Lengend Snippet: Intracellular cholesterol is increased via LDL or 25-HC treatment and decreased via simvastatin treatment . ( A ) Representative immunofluorescent filipin stain of cholesterol (purple) and pORF2 (green) upon treatment in persistently HEV-infected A549 cells; scale bar = 20 μm. ( B ) Quantification of A ; filipin intensity depicted as CTCF per cell. ( C ) qPCR evaluation of transcriptional effects of treatment with 25-HC in Huh-7 or A549/D3 cells, values referred to respective untreated control. ( D ) Cytotoxicity in persistently HEV-infected cells treated with different compounds as determined via lactate dehydrogenase assay; untreated cells were set to 100%, changes in amount of intact cells are depicted as % of untreated control. ( E ) Cytostaticity in persistently HEV-infected cells treated with different compounds as determined via PrestoBlue assay; untreated cells were set to 100%, changes in metabolic activity are depicted as % of untreated control. Unpaired t test with Holm-Sidak correction for all panels. ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, ∗∗∗∗ P < .0001. DMSO, dimethyl sulfoxide.

Article Snippet: Treatments with LDL (360-10-0.1; Lee Biosolutions, Maryland Heights, MO), 25-HC (11097; Cayman Chemicals, Ann Arbor, MI) and simvastatin (sc-200829B; Santa Cruz Biotechnology, Heidelberg, Germany) were performed in serum-free DMEM (D6546-500ML; Sigma Aldrich, Schnelldorf, Germany).

Techniques: Staining, Infection, Control, Lactate Dehydrogenase Assay, Prestoblue Assay, Activity Assay

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Targeting Cholesterol Metabolism as Efficient Antiviral Strategy Against the Hepatitis E Virus

doi: 10.1016/j.jcmgh.2021.02.002

Figure Lengend Snippet: HEV release is inhibited by LDL and 25-HC but induced by simvastatin. ( A ) Representative Western blot of pORF2 and GAPDH; neg = uninfected A549/D3 cells; black arrows indicate pORF2 bands. ( B–D ) Quantification of pORF2-signals in A ; fold-change compared with untreated group; unpaired t test with Holm-Sidak correction. ( E ) Fold-change of intracellular HEV transcripts as determined by RT-qPCR; unpaired t test with Holm-Sidak correction. ( F ) Fold-change of extracellular HEV RNA as determined by RT-qPCR; unpaired t test with Holm-Sidak correction. ( G ) Fold-change of released infectious viral particles as determined by end-point dilution assay; unpaired t test with Holm-Sidak correction. ( H ) HEV RNA in fractions of density-gradients as determined by RT-qPCR; depicted as % of whole genomes in gradient. ( I ) HEV titer in chronically infected patients with or without statin treatment; n = 42 patients, 129 measured values (without = 74 values; with = 55 values); Mann-Whitney-test. ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, ∗∗∗∗ P < .0001.

Article Snippet: Treatments with LDL (360-10-0.1; Lee Biosolutions, Maryland Heights, MO), 25-HC (11097; Cayman Chemicals, Ann Arbor, MI) and simvastatin (sc-200829B; Santa Cruz Biotechnology, Heidelberg, Germany) were performed in serum-free DMEM (D6546-500ML; Sigma Aldrich, Schnelldorf, Germany).

Techniques: Western Blot, Quantitative RT-PCR, End-point Dilution Assay, Infection, MANN-WHITNEY

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Targeting Cholesterol Metabolism as Efficient Antiviral Strategy Against the Hepatitis E Virus

doi: 10.1016/j.jcmgh.2021.02.002

Figure Lengend Snippet: Summary of Cholesterol- and Drug-Based Effects on HEV

Article Snippet: Treatments with LDL (360-10-0.1; Lee Biosolutions, Maryland Heights, MO), 25-HC (11097; Cayman Chemicals, Ann Arbor, MI) and simvastatin (sc-200829B; Santa Cruz Biotechnology, Heidelberg, Germany) were performed in serum-free DMEM (D6546-500ML; Sigma Aldrich, Schnelldorf, Germany).

Techniques:

Journal: PLoS ONE

Article Title: Comparative pharmacokinetics and safety assessment of transdermal berberine and dihydroberberine

doi: 10.1371/journal.pone.0194979

Figure Lengend Snippet: Precision and accuracy of berberine, simvastatin and simvastatin hydroxy acid in rat serum (inter-day n = 6x3; intra-day n = 6).

Article Snippet: Simvastatin (SIM, S485000), simvastatin hydroxy acid (SHA, S485020), d 6 -simvastatin (d6-SIM, S485002), d 6 -simvastatin hydroxy acid (d6-SHA, S485022), and d 6 -berberine hydrochloride (d6-BBR, B318152) were purchased from Toronto Research Chemicals.

Techniques: Concentration Assay

Journal: PLoS ONE

Article Title: Comparative pharmacokinetics and safety assessment of transdermal berberine and dihydroberberine

doi: 10.1371/journal.pone.0194979

Figure Lengend Snippet: Extraction recovery and matrix effect of BBR, SIM and SHA (n = 6) and d6-BBR, d6-SIM and d6-SHA (n = 6x3).

Article Snippet: Simvastatin (SIM, S485000), simvastatin hydroxy acid (SHA, S485020), d 6 -simvastatin (d6-SIM, S485002), d 6 -simvastatin hydroxy acid (d6-SHA, S485022), and d 6 -berberine hydrochloride (d6-BBR, B318152) were purchased from Toronto Research Chemicals.

Techniques: Concentration Assay

Journal: PLoS ONE

Article Title: Comparative pharmacokinetics and safety assessment of transdermal berberine and dihydroberberine

doi: 10.1371/journal.pone.0194979

Figure Lengend Snippet: Male Sprague-Dawley rats (N = 4/group) received once daily for 16 days administration of 90 mg/kg of active via BBR oral gavage (BBR PO, designated with green ο ), 5% (w/w) BBR transdermal formulation (BBR TD, designated with red ο ), 5% (w/w) DHB transdermal formulation (DHB TD; designated with blue ο ), or vehicle control (vehicle-TD, designated with black ο ). On day 16 all animals also received 12 mg/kg simvastatin and serum was collected over the course of eight hours. Concentrations of (A) SIM and (B) SHA were quantified. No statistical difference between groups was observed . Error bars represent standard error of the mean.

Article Snippet: Simvastatin (SIM, S485000), simvastatin hydroxy acid (SHA, S485020), d 6 -simvastatin (d6-SIM, S485002), d 6 -simvastatin hydroxy acid (d6-SHA, S485022), and d 6 -berberine hydrochloride (d6-BBR, B318152) were purchased from Toronto Research Chemicals.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Effect of Arthrospira maxima Phycobiliproteins, Rosiglitazone, and 17β-Estradiol on Lipogenic and Inflammatory Gene Expression during 3T3-L1 Preadipocyte Cell Differentiation

doi: 10.3390/ijms25147566

Figure Lengend Snippet: Effect of phycobiliproteins (PBPs) from A. maxima and simvastatin (SIM) on the viability of 3T3-L1 preadipocytes at 24 h. Values are shown as mean ± standard deviation (n = 12). Data were compared using a one-way ANOVA analysis with Tukey’s multiple comparison test. Asterisks indicate significantly different means (*** p < 0.001 and **** p < 0.0001).

Article Snippet: RSG was obtained from Cayman Chemical Co. (Ann Arbor, MI, USA), and simvastatin was obtained from Stress Marq Biosciences Inc. (Victoria, BC, Canada).

Techniques: Standard Deviation, Comparison

Journal: International Journal of Analytical Chemistry

Article Title: Comprehensive Assessment of Degradation Behavior of Simvastatin by UHPLC/MS Method, Employing Experimental Design Methodology

doi: 10.1155/2018/7170539

Figure Lengend Snippet: Obtained results from validation of the proposed method : robustness.

Article Snippet: Simvastatin CRS (purity 99.7% “as is”), Simvastatin impurity E (Lovastatin), and Simvastatin for peak identification were provided by European Directorate for the Quality of Medicines and Health Care Council of Europe (EDQM-Strasbourg, France).

Techniques: Biomarker Discovery

Journal: JCI insight

Article Title: Age-related decline in hippocampal tyrosine phosphatase PTPRO is a mechanistic factor in chemotherapy-related cognitive impairment.

doi: 10.1172/jci.insight.166306

Figure Lengend Snippet: Figure 4. Ptpro–/– female mice treated with DOX exhibit more severe neurodegeneration and impaired neurogenesis in the hippocampi following DOX treatment. (A) Nissl staining of the hippocampi (upper panel) and hippocampal CA3 region (bottom panel) in Ptpro+/+ and Ptpro–/– mice. Scale bars: 200 μm (upper panel), 50 μm (bottom panel). (B) Quantification of surviving neurons in the hippocampal CA3 region of Ptpro+/+ and Ptpro–/– mice. n = 5 mice per geno- type. (C) TUNEL staining of the hippocampi (upper panel) and hippocampal CA3 region (bottom panel) in Ptpro+/+ and Ptpro–/– mice. Scale bars: 250 μm (upper panel), 100 μm (bottom panel). (D) Quantification of TUNEL-positive hippo- campal CA3 cells in Ptpro+/+ and Ptpro–/– mice. n = 6 mice per genotype. (E) Representative images of immature (DCX+) and proliferating (Ki67+) cells in the hippocampi. Magnified views of areas in the white box are shown in the bottom left corner of each image. Scale bars: 100 μm. (F) Quantification of proliferating immature cells in the hippocampi. DG, dentate gyrus. n = 6 mice per genotype. These results are representative of 3 independent experiments. Error bars: SEM. ***P < 0.001 by 2-sided Student’s t test.

Article Snippet: For immunofluorescent staining, sections were incubated with antibodies against the following proteins: Ki67 (1:400; catalog 9129, Cell Signaling Technology), DCX (1:200; catalog sc-217390, Santa Cruz Biotechnology), PSD95 (1:200; catalog 3450, Cell Signaling Technology), and Syp (1:250; catalog ab32127, Abcam).

Techniques: Staining, TUNEL Assay

Journal: JCI insight

Article Title: Age-related decline in hippocampal tyrosine phosphatase PTPRO is a mechanistic factor in chemotherapy-related cognitive impairment.

doi: 10.1172/jci.insight.166306

Figure Lengend Snippet: Figure 8. Region-specific restoration of hippocampal PTPRO in Ptpro–/– female mice treated with DOX prevents neurodegeneration and promotes neurogenesis. (A) Nissl staining of the hippocampi (upper panel) and hippocampal CA3 region (bottom panel) in Ptpro+/+-LVCon, Ptpro+/+-LVPtpro, Ptpro–/–- LVCon, and Ptpro–/–-LVPtpro mice. Scale bars: 200 μm (upper panel), 50 μm (bottom panel). (B) Quantification of surviving neurons in the hippocampal CA3 region of Ptpro+/+-LVCon, Ptpro+/+-LVPtpro, Ptpro–/–-LVCon, and Ptpro–/–-LVPtpro mice. n = 5 per genotype. (C) TUNEL staining of the hippocampi (upper panel) and hippocampal CA3 region (bottom panel) in Ptpro+/+-LVCon, Ptpro+/+-LVPtpro, Ptpro–/–-LVCon, and Ptpro–/–-LVPtpro mice. Scale bars: 250 μm (upper panel), 100 μm (bottom panel). (D) Quantification of TUNEL-positive hippocampal CA3 cells in Ptpro+/+-LVCon, Ptpro+/+-LVPtpro, Ptpro–/–-LVCon, and Ptpro–/–-LVPtpro mice. n = 6 per genotype. (E) Representative images of immature (DCX+) and proliferating (Ki67+) cells in the hippocampi. Magnified views of areas in the white box are shown in the bottom left corner of each image. Scale bars: 100 μm. (F) Quantification of proliferating immature cells in the hippocampi. n = 6 per genotype. DG, dentate gyrus. These results are representative of 3 independent experiments. Error bars: SEM. NS, not signifi- cant; **P < 0.01, ***P < 0.001 by 1-way ANOVA followed by a Tukey-Kramer post hoc test.

Article Snippet: For immunofluorescent staining, sections were incubated with antibodies against the following proteins: Ki67 (1:400; catalog 9129, Cell Signaling Technology), DCX (1:200; catalog sc-217390, Santa Cruz Biotechnology), PSD95 (1:200; catalog 3450, Cell Signaling Technology), and Syp (1:250; catalog ab32127, Abcam).

Techniques: Staining, TUNEL Assay