Journal: Journal of neuropathology and experimental neurology

Article Title: Hereditary ferritinopathy: a novel mutation, its cellular pathology, and pathogenetic insights.

doi: 10.1093/jnen/64.4.280

Figure Lengend Snippet: FIGURE 9. Hyaline deposits. Pallidum. (A) Pleomorphic hyaline deposits, some containing nuclei, within the same area of GPe as in Figure 6A. (B) Prussian-blue reactivity. (C) PTAH reactivity. (D) Polyclonal anti-ferritin immunoreactivity. (E) Rare association of hyaline deposits with GFAP-immunoreactive astrocyte. (F) HO-2-immunoreactivity. Original magnifications: (A) Hematoxylin and eosin, 1503; (B) Perl’s stain, 2203; (C) 2203; (D) 2503; (E) 2203; (F) 2503.

Article Snippet: In order to identify the cell(s) of origin of the unique vacuolated nuclei, antibodies to S-100 (polyclonal, 1:20k, Lot #941; Innovation Foundation, Toronto, Ontario, Canada), GFAP, CD68 (monoclonal, 1:200, Lot #120201; DAKO), and carbonic anhydrase II (CAII) (polyclonal, 100-401-136/4383, 1:15k with retrieval; Rockland, Gilbertsville, PA) were applied to sections of basal ganglia and cerebellum, while antibodies to synaptophysin (monoclonal, 1:100 with retrieval, #M0363-UC/0403; Biogenex, San Ramon, CA), PGP 9.5 (polyclonal, 1:1,000 with retrieval, Lot #11830; Biogenesis, Handsown, NH), neurofilament protein 2F11 (monoclonal, 1:350, #M0762/117 DAKO) (101), and amyloid precursor protein (APP A4; monoclonal, 1:20K with heat retrieval, #23080547, Chemicon) were applied to the section of basal ganglia.

Techniques: Staining

Journal: Nature Communications

Article Title: Targeting leucine-rich repeat kinase 2 overcomes resistance to oncolytic herpes simplex virus-based therapies in glioblastoma

doi: 10.1038/s41467-026-70132-9

Figure Lengend Snippet: a Western blot analysis of LRRK2 and phosphorylated Rab10 (p-Rab10) protein expression in different GBM cell lines. The samples derive from the same experiment, but different gels for LRRK2, another for p-Rab10 and another for GAPDH were processed in parallel. b EC 50 values were determined in GBM cell lines treated with escalating titers of virus, with or without LRRK2-IN-1 (2.5 μM), for 72 h. c Pearson correlation analysis between the EC 50 values for oHSV-GFP and relative expression levels of LRRK2 or p-Rab10 in different GBM cell lines. Quantified LRRK2 expression was derived from band densities shown in Fig. 8a. d–g Effects of LRRK2-IN-1 on oHSV-GFP-mediated cytotoxicity in various normal cell types. EC 50 shifts were determined in primary astrocytes ( d ) human PBMCs-derived macrophages ( e ) iPSCs-derived neurons ( f ) or human microglia cells ( g ) treated with escalating titers of virus with or without LRRK2-IN-1 (2.5 μM), for 48 h. h Western blot analysis of LRRK2 protein expression in two GBM PDX tumors. The samples derive from the same experiment, but different gels for LRRK2 and another for actin were processed in parallel. i Experimental timeline for the PDX mouse model. NOD/SCID mice were subcutaneously inoculated with GBM PDX tumors. Once the tumor volume reached approximately 100–150 mm 3 , mice were treated with vehicle, oHSV-GFP virus (1 × 10 7 PFU, intratumorally every 2 days for a total of three injections), LRRK2-IN-1 (25 mg/kg, intraperitoneally every 2 days for a total of three injections), or a combination (Combo). j , k Tumor growth was monitored over time in GBM-PDX-1 ( n = 7) and GBM-PDX-2 ( n = 5). Western blot analyses were independently repeated three times for panel a, and twice for panel h, with similar results. The data are presented as the mean ± SD from three independent experiments ( b , d – g ), and P values were calculated using Spearman’s correlation test ( c ) and repeated measures ANOVA ( j , k ).

Article Snippet: Human astrocytes were purchased from iXCell Biotechnologies and cultured in astrocyte medium according to the manufacturer’s instructions.

Techniques: Western Blot, Expressing, Virus, Derivative Assay

Journal: Pharmaceuticals

Article Title: Transcriptomic Analysis of Fumarate Compounds Identifies Unique Effects of Isosorbide Di-(Methyl Fumarate) on NRF2, NF-kappaB and IRF1 Pathway Genes

doi: 10.3390/ph15040461

Figure Lengend Snippet: RT-PCR analyses and NF-κB luciferase reporter assay. ( A – G ) RT-PCR analyses ( OSGIN1 , IL6 , ICAM1 , MALT1 , TNFAIP3 , IRF1, and CXCL8 ). Average relative expression is shown for each gene (±1 standard error; n = 3 replicates per treatment). Relative expression was calculated using the 2 −∆∆Ct method and ∆Ct values were further normalized to the CTL treatment. Average relative expression is shown for each treatment (bottom margin). Treatments with different letters have significantly different average expression ( p < 0.05, Fisher’s least significant difference). ( H ) NF-κB reporter assay. Human fetal astrocytes were transfected with firefly/Renilla luciferase constructs and treated with TNF-α (20 ng/ml) alone ( n = 6) or TNF-α (20 ng/mL) + IDMF (2.5 µM) ( n = 3) for 6 h. The ratio of luciferase and Renilla (internal control) signals was used to monitor NF-κB induction. The average ratio value is shown (±1 standard error) for each treatment ( p -value: two-sample Wilcoxon rank sum test).

Article Snippet: Experiments were performed using human fetal astrocytes (HA) derived from cerebral cortex (cat no. 882A-05f; Cell Applications, San Diego, CA, USA) grown to subconfluence for 48 h in HA growth medium (cat. no. 820-500, Cell Applications) in a 96-well black wall tissue culture plate.

Techniques: Reverse Transcription Polymerase Chain Reaction, Luciferase, Reporter Assay, Expressing, Transfection, Construct, Control