Journal: bioRxiv

Article Title: RAD52 prevents accumulation of Polα-dependent replication gaps at perturbed replication forks in human cells

doi: 10.1101/2023.04.12.536536

Figure Lengend Snippet: A. Analysis of Polα-RAD51 association by PLA in U2OS WT. Cells were subjected to RAD52i 30 min before HU (2 mM) treatment. PLA reaction was carried out using antibodies against POLA1 and RAD51. The graph shows the number of PLA spot per nucleus. The control was subjected to PLA with only one primary antibody. Representative images are shown. Scale bar represents 10 μm B. Analysis of Polα-RAD51 association by PLA in U2OS WT. Cells were exposed to RAD52i 20 min before HU (0.5 mM) treatment of 2 and 4 hours. PLA reaction was carried out using antibodies against POLA1 and RAD51. The graph shows the number of PLA spot per nucleus. C. Experimental scheme of super-resolution (SR) microscopy assay. D. Quantification of EdU-RAD51-Polα interaction by SR microscopy (dSTORM). U2OS WT were treated as indicated on the panel C. The graph represents the percentage of variation in co-localization events of EdU-RAD51, EdU- Polα and EdU-RAD51-Polα. E. Representative dSTORM images of two nuclei immunolabeled for nascent DNA (cyan), Polα (magenta) and RAD51 (yellow). Scale bars = 20μm, 1μm and 100nm. A representative sketch of a common topology of the 3 signal is shown. All the values above are presented as means ± SE (ns = not significant; *P < 0.1; **P<0.1; ***P < 0.001; ****P < 0.0001; Mann–Whitney test;).

Article Snippet: The fixed cells on coverslips were mounted onto concave slides and dSTORM imaging B3 buffer from Oxford Nanoimaging (ONI, PN #900–00004) was added before imaging.

Techniques: Control, Microscopy, Immunolabeling, MANN-WHITNEY

Journal: iScience

Article Title: Identification of IQCH as a calmodulin-associated protein required for sperm motility in humans

doi: 10.1016/j.isci.2023.107354

Figure Lengend Snippet: Analysis of the spatial distribution of IQCH within sperm flagella by super-resolution microscopy (A) STORM imaging of sperm flagella stained for IQCH (red) and α-tubulin (green) or calmodulin (CaM, green). Scale bars 1 μm. (B) STED images of co-staining IQCH (violet) and α-tubulin (green), highlighting the helicoidal spatial distribution of IQCH along the sperm flagellum (dashed frames) and its co-localization with calmodulin. Distribution profiles of the signal in the selected regions highlight the longitudinal and transversal periodicity of IQCH. Scale bars 2 μm and 2.5 μm. (C) Schematic representation of a transversal section of a sperm flagellum with the localization of proteins identifying the various complexes and subcellular regions of sperm flagella localization [Graphical elements from Servier Medical Art, licensed under a Creative Commons Attribution 3.0 Unported License]. Distribution data were obtained by analyzing the STORM and STED images shown in Figure S4 .

Article Snippet: Mounting buffer kit for STORM imaging , Abbelight , Smart kit.

Techniques: Super-Resolution Microscopy, Imaging, Staining

Journal: iScience

Article Title: Identification of IQCH as a calmodulin-associated protein required for sperm motility in humans

doi: 10.1016/j.isci.2023.107354

Figure Lengend Snippet:

Article Snippet: Mounting buffer kit for STORM imaging , Abbelight , Smart kit.

Techniques: Control, Recombinant, Protease Inhibitor, Electron Microscopy, Plasmid Preparation, Reverse Transcription, Gel Extraction, SYBR Green Assay, Imaging, In Situ, Software