Journal: Journal of Alzheimer's Disease

Article Title: Dihydropyridine Derivatives Modulate Heat Shock Responses and have a Neuroprotective Effect in a Transgenic Mouse Model of Alzheimer’s Disease

doi: 10.3233/jad-150860

Figure Lengend Snippet: Fig. 3. Effect of heat and LA1011 or LA1044 treatment on Hsp70, Hsp27 and Hsp40 levels in SH-SY5Y human neuroblastoma cell line measured by western blotting (A, B) and quantitative densitometric analysis (C, D). Cells were untreated or treated with LA1011 (5 M) or LA1044 (40 M) at 37◦C or 42◦C. Box plots represent quantitative data normalized to GAPDH. All data represent the mean ± SD (n = 6) and p < 0.05 was considered statistically significant. HS = heat shock at 42◦C, 1 h. ∗p < 0.05, when data are compared to LA compound (–), HS (–) sample. #p < 0.05, when data are compared to LA compound (–), HS (+) sample. One way ANOVA followed by a Tukey post hoc test was used for statistical comparisons.

Article Snippet: The effect of drug candidates on the stress response was tested with the SH-SY5Y (ATCC® CRL-2266TM) human neuroblastoma cell line stably transfected with a plasmid containing the Hsp70 promoter fused to yellow fluorescent protein (YFP) (pEYFP-N1, Clontech), a kind gift of Lea Sistonen (Åbo Akademi University, Turku, Finland) (SH-SY5Y-pHsp70-YFP).

Techniques: Western Blot

Journal: Nature Communications

Article Title: RiboBright reveals cell-type-specific differences in ribosome organization and movement

doi: 10.1038/s41467-026-68947-7

Figure Lengend Snippet: a Representative single z-plane images of mESCs, HCT 116, HEK293T/17, MCF 10A, SH-SY5Y, PC-9, HeLa, U2OS, SK-MEL-28, and PANC−1 cells stained with 10 μM RiboBright. Images were acquired with 100x/1.40 NA oil objective ( n = 2 biological replicates). Scale bars are 15 μm, 1 μm. b Distributions of flow cytometry data show live mESCs, HCT 116, HEK293T/17, MCF 10 A, SH-SY5Y, PC-9, HeLa, U2OS, SK-MEL-28, and PANC-1 cells stained with 10 μM RiboBright. The unstained control is subtracted for each cell type. c Representative single z-plane images of fixed HEK293T/17 cells co-stained with OPP and RiboBright. Images were acquired with 100x/1.40 NA oil objective ( n = 2 biological replicates). Scale bars are 10 μm. d Scatter plot showing the relationship between OPP intensity (i.e., translation levels) and RiboBright intensity (i.e., ribosome content) measured in single cells in mESCs (400 cells), HCT 116 (557 cells), HEK293T/17 (618 cells), MCF 10 A (657 cells), SH-SY5Y (402 cells), PC-9 (284 cells), HeLa (498 cells), U2OS (502 cells), SK-MEL-28 (339 cells) and PANC-1 (712 cells) cells within one biological replicate. Each data point corresponds to an individual cell per cell type. e Pearson correlation between the RiboBright intensities and variability (measured as Fano factor = σ 2 /μ) across the cell lines shown in ( d ) and Supplementary Fig . f Average per-ribosome translational efficiency measured as the ratio between OPP and RiboBright intensity (=OPP (new protein synthesis)/RiboBright (Ribosome content)) measured across the cell lines shown in ( d ) and Supplementary Fig. . e , f Data was acquired for 888 mES cells, 2132 HCT 116 cells, 1338 HEK293T/17 cells, 1412 MCF 10 A cells, 755 SH-SY5Y cells, 730 PC-9 cells, 1165 HeLa cells, 1041 U2OS cells, 941c SK-MEL-28 cells and 1675 PANC-1 cells, each dot indicates the mean of the two biological replicates ( n = 2).

Article Snippet: HEK293T/17 (ATCC), SH-SY5Y (Prof. Ger Pruijn, Radboud University), HeLa (Prof. Wilhelm Huck, Radboud University), U2OS (Dr. Klaas Mulder, Radboud University), PANC-1 (Prof. Rene Bernard, Netherlands Cancer Institute) and HCT 116 cells (Dr. Klaas Mulder, Radboud University) were cultured in Dulbecco’s modified Eagle’s medium (DMEM, ThermoFisher) supplemented with 4.5 g/L D-Glucose, L-glutamine, Sodium Pyruvate, 10% (v/v) fetal bovine serum (FBS, ThermoFisher) and antibiotic solution (50 U/mL Penicillin and 50 U/mL Streptomycin). mESC-E14 (mESCs) (129/Ola background) were obtained from Dr. Hendrik Marks, originally from ATCC, with RRID:CVCL_9108 .

Techniques: Staining, Flow Cytometry, Control