Journal: FEBS Open Bio

Article Title: Differential heme release from various hemoglobin redox states and the upregulation of cellular heme oxygenase‐1

doi: 10.1002/2211-5463.12103

Figure Lengend Snippet: Hemoglobin‐mediated induction of heme oxygenase‐1 ( HO ‐1) expressions in mouse E10 cells. E10 cells were exposed to either ferrous, ferric, or ferryl Hb at equimolar concentration (100 µ m ) for 12 h for immunocytochemistry experiment (A) or 24 h for western blotting experiment (B). Laser confocal images showing expression of heme oxygenase‐1 (green, Alexa‐488) in E10 cells (A). E10 cells were also counterstained with Phalloidin Alexa‐647 (red) to show the cellular morphology. Nuclear staining was done with DAPI (blue). Cell lysates were immunoblotted and analyzed for HO ‐1 expression (B, upper panel). Equal loading was confirmed by reprobing the blots against β‐actin (B, lower panel). The ratio of average ( n = 3) band intensity of HO ‐1 with corresponding β‐actin was plotted as relative intensity (C), * P < 0.05 vs. control; # P < 0.05 vs. HbFe 3+ , ‘ t ’ test unpaired.

Article Snippet: Cells were visualized under a Zeiss LSM710 meta confocal microscope (Zeiss, Thornwood, NY, USA) after mounting with Fluoroshield mounting medium with DAPI (Abcam).

Techniques: Concentration Assay, Immunocytochemistry, Western Blot, Expressing, Staining, Control

Journal: Human Genetics

Article Title: De novo mutations in FBRSL1 cause a novel recognizable malformation and intellectual disability syndrome

doi: 10.1007/s00439-020-02175-x

Figure Lengend Snippet: Immunofluorescence analysis performed on HEK293 cells ( a ) and human fibroblasts ( b ). The N-terminal antibody detected isoforms 1, 3.1 and 3.2 in the cytoplasm and nucleus. Interestingly, an association with centrosomes (white arrow) and kinetochores was detected. Staining with a C-terminal antibody, detecting the full-length isoform 1 and additional hypothetic short C-terminal isoforms, showed a mainly nuclear pattern without a co-localization with the mitotic spindle, centrosomes or kinetochores. α-Tubulin was used for cytoskeletal staining and nuclei were stained using DAPI. Images were obtained using a confocal laser microscope with × 600 magnification, and an additional software magnification as indicated in the respectively images

Article Snippet: Subsequently, cells were washed twice and mounted using FluoroshieldTM with DAPI (Merck Millipore).

Techniques: Immunofluorescence, Staining, Microscopy, Software

Journal: Nature Communications

Article Title: Integration of tumor extrinsic and intrinsic features associates with immunotherapy response in non-small cell lung cancer

doi: 10.1038/s41467-022-31769-4

Figure Lengend Snippet: UMAP projections showing the expression of a HLA-I and HLA-II in the CD45 − fraction. HLA-I and II expression was measured by summing the expression of all the individual genes assessed in the locus. b Comparison of HLA-II and CD74 (invariant chain) expression ( R = 0.751, unadjusted p = 0, Pearson correlation test). c Boxplots show the log-transformed expression of the individual HLA-II genes assessed ( p = 1.01 × 10 −178 , Kruskal–Wallis, n = 1377 cells). The box represents the interquartile range, with the center line at the median. The whiskers extend up to 1.5 times the interquartile range (IQR). d Representative multiplex immunofluorescent staining of CD8 (blue), CD4 (red), GZMB (green), PanCK (magenta), HLA-DR (cyan), and DAPI (gray) in an NSCLC tumor ( n = 2). Overlay without DAPI is shown for CD8, CD4, and GZMB, and for PanCK and HLA-DR, along with the corresponding H&E. Scale bar, 25 μm. Source data are provided as a Source Data file.

Article Snippet: Slides were then mounted using Fluoroshield with DAPI (Immunobiosciences) and coverslips applied prior to scanning at 20× using the Leica Versa scanner.

Techniques: Expressing, Comparison, Transformation Assay, Multiplex Assay, Staining