Journal: Developmental biology

Article Title: Fertilization stimulates 8-hydroxy-2'-deoxyguanosine repair and antioxidant activity to prevent mutagenesis in the embryo.

doi: 10.1016/j.ydbio.2015.07.024

Figure Lengend Snippet: Fig. 1. OGG1 is under-represented in MII-stage mouse oocytes. (A) Quantitative PCR analysis revealed a low abundance of OGG1 and APE1 mRNA within the murine oocyte (relative to B2M and Gusβ), whilst XRCC1 transcripts were present at significantly higher levels. (B) Immunoblotting analysis of the protein composition of these oocytes identified bands corresponding to APE1 and XRCC1 at 35 kDa and 85 kDa respectively; whilst a 37 kDa band corresponding to OGG1 could not be detected at this con- centration of protein (100 oocytes per lane, equivalent to 1 mg protein). Positive controls; rOGG1, rAPE1 and Jurkat cell lysate respectively (1 mg). (C, D) In support of Western blotting data, immunocytochemistry identified fluorescence associated with APE1 and XRCC1 throughout the ooplasm. Fluorescence relating to OGG1 was low, however upon pixel intensity analysis was still found to be significantly greater than the ‘secondary only’ control (“C”). Scale bar¼50 mm. Mean7SEM values are plotted in histograms. Independent replicates were conducted with a minimum of 40 oocytes per replicate. *Po0.05, **Po0.01, ***Po0.001.

Article Snippet: 1 mg of recombinant OGG1 (rOGG1), recombinant APE1 (rAPE1) (both from New England Biolabs, Ipswich, MA, USA), and Jurkat whole cell lysate (Novus Biologicals, Littleton, CO, USA) was loaded onto the polyacrylamide gels as a positive control for OGG1, APE1 and XRCC1 antibodies respectively.

Techniques: Real-time Polymerase Chain Reaction, Western Blot, Immunocytochemistry, Fluorescence, Control

Journal: Frontiers in Immunology

Article Title: Deep immune-phenotyping of HLA-homozygous iPS-cardiomyocytes by spectral flow cytometry

doi: 10.3389/fimmu.2026.1736994

Figure Lengend Snippet: Stability of the iPS-CM immune-phenotype across different production sites. (A) Dimensionality reduction (tSNE) of 6 independent iPS-CM differentiations of HLA-homozygous R26 performed at 3 different production sites with and without final IFN-γ challenge. Separate depiction of marker combination in three spectral flow cytometry analysis tubes (t1-3) as shown. Color-coding represents sample identity. (B) Tube 2 and (C) tube 3 tSNE blots showing the expression heat of markers driving the separation of clusters in samples. (D) Heatmap showing the overall expression profiles of viable cells in samples from individual differentiation batches across all 54 markers plus cTNT. Marker profiles of Jurkat cells and PBMCs were included as a reference. Median fluorescence intensity (MFI) was scaled column-wise.

Article Snippet: Jurkat cell stocks were purchased from the German Collection of Microorganisms and Cell Cultures (DSMZ; Jurkat ACC282) and expanded in RPMI-1640 (Sigma-Aldrich) containing 10% FBS (Gibco), 5 mM N(2)-L-Alanyl-L-Glutamin (Dipeptiven, Fresenius Kabi), and 10 mM HEPES (Sigma-Aldrich) before cryopreservation in a working cell bank at -170°C until further use in experiments.

Techniques: Marker, Flow Cytometry, Expressing, Fluorescence

Journal: Cancer Cell International

Article Title: IGFBP3 induces PD-L1 expression to promote glioblastoma immune evasion

doi: 10.1186/s12935-024-03234-3

Figure Lengend Snippet: IGFBP3 induces immunosuppression in in-vitro co-culture system. A CCK-8 assay was used to analysis the viability of Jurkat cells after co-cultured with IGFBP3 overexpressing cells. B , C QRT-PCR was performed to examine the mRNA expression of IL2 ( B ) or IFN-γ ( C ) in Jurkat cells after co-cultured with IGFBP3 overexpressing cells. D , E ELISA was used to measure the levels of IL-2 ( D ) and IFN-γ ( E ) in the supernatants of Jurkat cells after co-cultured with IGFBP3 overexpressing cells. F Annexin V-PE apoptosis assay was used to determine apoptosis of Jurkat cells after co-cultured with IGFBP3 overexpressing cells. G Statistical chart of apoptotic cells. H Immunoblotting was performed to examine the caspase3 expression in Jurkat cells after co-cultured with IGFBP3 overexpressing cells. Data are expressed as mean ± SD. *P < 0.05; **P < 0.01; ***P < 0.001, Student t test

Article Snippet: Briefly, after co-culture with tumor cells, Jurkat cells were collected and stained with annexin V-FITC and PI (Elabscience, E-CK-A211).

Techniques: In Vitro, Co-Culture Assay, CCK-8 Assay, Cell Culture, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Apoptosis Assay, Western Blot