Journal: Scientific Reports

Article Title: Insight into the cellular fate and toxicity of aluminium adjuvants used in clinically approved human vaccinations

doi: 10.1038/srep31578

Figure Lengend Snippet: Representative lumogallion staining of agar-paraffin embedded (2 μm sections) THP-1 cells co-cultured with ( a ) 2.5, ( b ) 25.0, ( c ) 50.0 or ( d ) 100.0 μg/mL Alhydrogel ® (Brenntag Biosector, Denmark). Cell sections were incubated for 24 h in 100 μM lumogallion, 50 mM PIPES, pH 7.4. Slides were mounted with ProLong ® Gold Antifade Reagent with DAPI. All images depict lumogallion staining (orange) overlaid with DAPI-staining (blue). Magnified inserts show close-ups of individual cells with the light channel overlaid. White arrows highlight both individual and distinguishable adjuvant particles. Magnification X 1000, scale bars: 20 μm.

Article Snippet: Sections were finally air dried and mounted using ProLong ® Gold Antifade Reagent with 4′,6-diamidino-2-phenylindole, dihydrochloride (DAPI) (Life Technologies, UK) prior to storing horizontally for 24 h at 4 °C.

Techniques: Staining, Cell Culture, Incubation, Adjuvant

Journal: Scientific Reports

Article Title: Insight into the cellular fate and toxicity of aluminium adjuvants used in clinically approved human vaccinations

doi: 10.1038/srep31578

Figure Lengend Snippet: Representative lumogallion staining of agar-paraffin embedded (2 μm sections) THP-1 cells co-cultured with ( a ) 2.5, ( b ) 25, ( c ) 50 or ( d ) 100 μg/mL Adju-Phos ® (Brenntag Biosector, Denmark). Cell sections were incubated for 24 h in 100 μM lumogallion, 50 mM PIPES, pH 7.4. Slides were mounted with ProLong ® Gold Antifade Reagent with DAPI. All images depict lumogallion staining (orange) overlaid with DAPI-staining (blue). Magnified inserts show close-ups of individual cells with the light channel overlaid. White arrows highlight both individual and distinguishable adjuvant particles. Magnification X 1000, scale bars: 20 μm.

Article Snippet: Sections were finally air dried and mounted using ProLong ® Gold Antifade Reagent with 4′,6-diamidino-2-phenylindole, dihydrochloride (DAPI) (Life Technologies, UK) prior to storing horizontally for 24 h at 4 °C.

Techniques: Staining, Cell Culture, Incubation, Adjuvant

Journal: Scientific Reports

Article Title: Insight into the cellular fate and toxicity of aluminium adjuvants used in clinically approved human vaccinations

doi: 10.1038/srep31578

Figure Lengend Snippet: Representative lumogallion staining of agar-paraffin embedded (2 μm sections) THP-1 cells co-cultured with ( a ) 2.5, ( b ) 25, ( c ) 50 or ( d ) 100 μg/mL Imject™ Alum (Thermo Scientific). Cell sections were incubated for 24 h in 100 μM lumogallion, 50 mM PIPES, pH 7.4. Slides were mounted with ProLong ® Gold Antifade Reagent with DAPI. All images depict lumogallion staining (orange) overlaid with DAPI-staining (blue). Magnified inserts show close-ups of individual cells with the light channel overlaid. White arrows highlight both individual and distinguishable adjuvant particles. Magnification X 1000, scale bars: 20 μm.

Article Snippet: Sections were finally air dried and mounted using ProLong ® Gold Antifade Reagent with 4′,6-diamidino-2-phenylindole, dihydrochloride (DAPI) (Life Technologies, UK) prior to storing horizontally for 24 h at 4 °C.

Techniques: Staining, Cell Culture, Incubation, Adjuvant

Journal: Oncotarget

Article Title: Silencing of casein kinase 1 delta reduces migration and metastasis of triple negative breast cancer cells

doi: 10.18632/oncotarget.25738

Figure Lengend Snippet: ( A ) Scatter plot of gene expression analysis by RT 2 Profiler PCR Array in the shNT MDA-MB-231 cells compared to shCSNK1D cells. The central line indicates unchanged gene expression. ( B ) Taqman validation of genes identified in the RT Profiler array. The histogram represents fold change of genes up-or down-regulated in shCSNK1D compared to shNT control cells. ( C ) Western blot analysis of the expression of occludin (OCLN), junction adhesion molecule A (F11R) and claudin-1 (CLDN-1) in control (shNT) and inactivated cell lines (shCSNK1D). ( D ) Detection of OCLN and F11R by immunofluorescence in shCSNK1D compared to control shNT cells. Nuclei were stained with Dapi. Arrows indicate the increased expression of tight junction proteins.

Article Snippet: After washing, the cells were mounted with Prolong gold antifade medium with DAPI (Cell Signaling).

Techniques: Gene Expression, Biomarker Discovery, Control, Western Blot, Expressing, Immunofluorescence, Staining

Journal: Redox Biology

Article Title: Accumulated cholesterol protects tumours from elevated lipid peroxidation in the microenvironment

doi: 10.1016/j.redox.2023.102678

Figure Lengend Snippet: Depletion of TME cholesterol promotes ferroptosis and LPO in vivo. A. MβCD treatment reduces cholesterol content in xenografts. B. MβCD treatment depletes LDL cholesterol in the TIF, as analysed by fast protein liquid chromatography (monitored through absorbance at 280 nm). Cholesterol was measured by Amplex Red. C. Growth curve and tumour weight of A375 xenografts in BALB/c nude mice with the indicated treatments. The weight of xenografts was obtained at the end of the indicated treatments. Each data point represents an individual xenograft tumour. D. LDLR protein analysed by western blotting in A375 xenografts treated as indicated. E. MDA content measured in xenografts. Tumour masses were cut from 3 mice in each group, and each mouse donated 3–4 tissue blocks. Each data point represents an individual tissue block. F-G. Immunofluorescence staining and quantification of 4-HNE-modified proteins (green) in paraffin-embedded xenografts at the end of the indicated treatment. Blue indicates DAPI-stained nuclei. Scale bars = 50 μm. Statistical significance was assessed using two-way ANOVA ( C ) or an unpaired two-tailed t -test (C, E-F). **P < 0.01, ***P < 0.01, ****P < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Then, anti-fade mounting medium with DAPI (S2100, Solarbio) was mounted.

Techniques: In Vivo, Fast Protein Liquid Chromatography, Western Blot, Blocking Assay, Immunofluorescence, Staining, Modification, Two Tailed Test