Journal: Communications Biology

Article Title: Lysine malonylation regulates human sperm motility

doi: 10.1038/s42003-026-09683-y

Figure Lengend Snippet: A The Kmal in total proteins of (TP) and protein fractions of the mitochondrion (Mit), head, and cytoplasm (Cyt) isolated from human sperm were examined by Western blot. The specificity of cellular components was validated by Western blot using the corresponding markers: PRM2 for sperm head, COX6B1 for mitochondrion, and ACTIN for cytoplasm. The proteins for Western blot were shown in the coomassie brilliant blue (CBB) staining image. B , C The localization of malonylated proteins within human sperm was investigated by immunofluorescence assay via laser scanning confocal microscopy (LSCM, ( B )) and super-resolution structured illumination microscopy (SIM, ( C )). All the experiments were conducted with samples from 8 normozoospermic men. The scale bar represents 5 μm.

Article Snippet: Anti-ACTIN antibody (66009-1-Ig), anti-SIRT5 antibody (67257-1-Ig), anti-acetyl-CoA carboxylase 1 (ACC1) antibody (21923-1-AP), anti-fatty acid synthase (FASN) antibody (10624-2-AP), anti-protamine 2 (PRM2) antibody (14500-1-AP), anti-GAPDHS (83290-3-RR) and anti-VDAC3 (82666-14-RR) were obtained from Proteintech Group, Inc. (Rosemont, IL, USA).

Techniques: Isolation, Western Blot, Staining, Immunofluorescence, Confocal Microscopy, Microscopy

Journal: F&S science

Article Title: Blastocyst development after fertilization with in vitro spermatids derived from nonhuman primate embryonic stem cells

doi: 10.1016/j.xfss.2021.09.001

Figure Lengend Snippet: Nonhuman primate embryonic stem cell (nhpESC) histone 2B-green fluorescent protein (H2B-GFP) cells differentiated in spermatogonial stem cell (SSC) conditions yield haploid, round spermatid-like cells. ( A ) Representative cell cycle flow cytometry profile data from one of five separate differentiations (n = 5) ( left image ) of nhpESC H2B-GFP cells differentiated in SSC conditions for 10 days. The dark green peak on the left-hand side of the graph represents the 1N peak ( asterisk ). ( B ) Fluorescence in situ hybridization (FISH) confirms haploidy. Representative FISH images from five different (n = 5) nhpESC H2B-GFP cells differentiated for 10 days in SSC conditions. The FISH Probe for chromosome 1 ( green ) shows a diploid cell ( left image ) with two chromosome 1s ( arrow indicating two green dots ), and sorted haploid cells ( right image ) show one chromosome 1 (arrow indicating one green dot ). Bar = 5 μ m. ( C ) Following our 10-day differentiation, haploid cells were sorted by fluorescence-activated cell sorting and immunostained with antibodies against postmeiotic spermatid markers ( red , third column ) acrosin ( top row ), Protamine 1 (PRM1; middle row ), and Transition Protein 1 (TNP1; bottom row ). H2B-GFP ( green , second column ); Hoechst stain ( blue , first column ) for deoxyribonucleic acid. Merge of fluorescence images ( fourth column ). Bar = 10 μ m. Data shown are representative of five separate differentiations (n = 5). ( D ) Representative images of round spermatids from a rhesus testis cell biopsy ( left ) and an in vitro-derived round spermatid-like cell ( right ) stained with 4′,6-diamidino-2-phenylindole ( blue ), Protamine 1 ( red ), and acetylated tubulin ( green ). Bar = 5 μ m.

Article Snippet: Acrosin (ACR), Protamine 1 (PRM1), and Transition Protein 1 were from Santa Cruz Biotechnology.

Techniques: Flow Cytometry, Fluorescence, In Situ Hybridization, FACS, Staining, In Vitro, Derivative Assay