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: Molecular Therapy. Nucleic Acids

Article Title: Distinct Expression Profiles and Novel Targets of MicroRNAs in Human Spermatogonia, Pachytene Spermatocytes, and Round Spermatids between OA Patients and NOA Patients

doi: 10.1016/j.omtn.2017.09.007

Figure Lengend Snippet: Morphological and Phenotypic Characterization of Freshly Isolated Human Spermatogonia, Pachytene Spermatocytes, and Round Spermatids (A–C) Phase-contrast microscope revealed the morphology of the freshly isolated human pachytene spermatocytes (A), spermatogonia (B), and round spermatids (C) of OA patients. (D–F) DIC microscope showed the morphological characteristics of the freshly isolated human pachytene spermatocytes (D), spermatogonia (E), and round spermatids (F) of OA patients. Scale bars, 20 μm (A–C) and 5 μm (D–F). (G) RT-PCR revealed the transcripts of GPR125 , RET , GFRA1 , THY1 , UCHL1 , MAGEA4 , and PLZF in the fleshly isolated spermatogonia, the expression of SYCP3 and SYCP1 in pachytene spermatocytes, and mRNA of TNP1 , TNP2 , PRM1 , PRM2 , and ACR in round spermatids. RNA without RT (RT-) but with PCR of GAPDH primers was utilized as negative controls, and GAPDH served as loading controls of total RNA.

Article Snippet: The primary antibodies included THY1 (Abcam, ab133350, 1:200), GFRA1 (Santa Cruz, sc-6156, 1:200), PLZF (Santa Cruz, sc-22839, 1:200), UCHL1 (Bio-Rad, MCA4750, 1:200), PNA (Life Technologies, L32458, 1:200), and Protamine 2 (PRM2) (Atlas Antibodies, HPA056386, 1:200).

Techniques: Isolation, Microscopy, Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Molecular Therapy. Nucleic Acids

Article Title: Distinct Expression Profiles and Novel Targets of MicroRNAs in Human Spermatogonia, Pachytene Spermatocytes, and Round Spermatids between OA Patients and NOA Patients

doi: 10.1016/j.omtn.2017.09.007

Figure Lengend Snippet: Phenotypic Characterization of Freshly Isolated Human Spermatogonia, Pachytene Spermatocytes, and Round Spermatids (A–D) Immunocytochemistry revealed the expression of THY1 (A), GFRA1 (B), PLZF (C), and UCHL1 (D) in the freshly isolated human spermatogonia. Scale bars, 10 μm. (E) Meiotic chromatin spread by triple immunostaining displayed the co-expression of CREST, SYCP3, and MLH1 in the freshly isolated human pachytene spermatocytes. Scale bar, 5 μm. (F–I) Immunocytochemistry demonstrated the expression of PNA (F), PRM2 (G), and rabbit IgG (I) in the freshly isolated human round spermatids and rabbit IgG (H) in the freshly isolated human spermatogonia. Scale bars, 10 μm.

Article Snippet: The primary antibodies included THY1 (Abcam, ab133350, 1:200), GFRA1 (Santa Cruz, sc-6156, 1:200), PLZF (Santa Cruz, sc-22839, 1:200), UCHL1 (Bio-Rad, MCA4750, 1:200), PNA (Life Technologies, L32458, 1:200), and Protamine 2 (PRM2) (Atlas Antibodies, HPA056386, 1:200).

Techniques: Isolation, Immunocytochemistry, Expressing, Triple Immunostaining

Journal: Molecular Therapy. Nucleic Acids

Article Title: Distinct Expression Profiles and Novel Targets of MicroRNAs in Human Spermatogonia, Pachytene Spermatocytes, and Round Spermatids between OA Patients and NOA Patients

doi: 10.1016/j.omtn.2017.09.007

Figure Lengend Snippet: The Sequences of Gene Primers Used for RT-PCR and Real-Time PCR

Article Snippet: The primary antibodies included THY1 (Abcam, ab133350, 1:200), GFRA1 (Santa Cruz, sc-6156, 1:200), PLZF (Santa Cruz, sc-22839, 1:200), UCHL1 (Bio-Rad, MCA4750, 1:200), PNA (Life Technologies, L32458, 1:200), and Protamine 2 (PRM2) (Atlas Antibodies, HPA056386, 1:200).

Techniques:

Journal: Communications Biology

Article Title: Lysine malonylation regulates human sperm motility

doi: 10.1038/s42003-026-09683-y

Figure Lengend Snippet: The levels of Kmal and SIRT5 were analyzed in 53 normozoospermic men (NOR) and 41 asthenozoospermic men (AST) by western blot. A An example illustrating the presence of sperm Kmal and SIRT5 in AST and NOR. B Semi-quantitative analysis of Kmal in NOR and AST was conducted as outlined in the “Materials and methods” section. The red box indicates 10 AST whose sperm Kmal levels exceeded 1.51, the highest value of relative Kmal levels in NOR. C Correlations between sperm Kmal and progressive motility in 94 samples were evaluated through linear regression analysis. The glycolysis ( D ) and ATP level ( E ) were examined in 10 AST indicated in the red box in Fig. 3B. F Semi-quantitative analysis of SIRT5 in NOR and AST was conducted as outlined in the “Materials and methods” section. G Correlations between sperm Kmal and SIRT5 in 94 samples were assessed through linear regression analysis. The error bars represent the means ± SEM. The differences between NOR and AST were analyzed by unpaired t-test with statistical significance indicated by * p < 0.05 and *** p < 0.001.

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: Western Blot