Journal: Scientific Reports

Article Title: Identification of ITPR1 gene as a novel target for hsa-miR-34b-5p in non-obstructive azoospermia: a Ca 2+ /apoptosis pathway cross-talk

doi: 10.1038/s41598-023-49155-5

Figure Lengend Snippet: Histological feature of the human testicular tissues. ( a–c ) H&E stained testis section from adult OA and NOA. ( a ) OA testicle (Control). ( b ) NOA testicle. ( c ) NC. Magnification: × 40. H&E Hematoxylin and eosin, NOA non-obstructive azoospermia, OA obstructive azoospermia, NC negative control. Arrows: Irregularity in NOA patients’ seminiferous. ( d–f ) TUNEL-stained sections of the human testicular tissue for the detection of apoptosis. ( d ) OA, control group. A few TUNEL-stained nuclei were observed in the OA (Arrowheads). ( e ) In the NOA, the increase in the apoptotic cells was observed compared to the OA group (Arrows). ( f ) NC negative control. Green and Red fluorescent stained nuclei indicate apoptotic and viable cells, respectively. Magnification, × 40. TUNEL deoxy-UTP-digoxigenin nick end labeling, NC negative control. ( g ) Quantification of staining intensity reveals 2.7-fold increases in apoptosis rate in NOA compared with OA (control group).

Article Snippet: Then, 5-μm-thick cross-sections were prepared for subsequent hematoxylin–eosin (H&E) staining and deoxy-UTP-digoxigenin nick end labeling (TUNEL) assay (Promega kit, USA).

Techniques: Staining, Control, Negative Control, TUNEL Assay, End Labeling

Journal: Transcription

Article Title: Activation and reactivation of the RNA polymerase II trigger loop for intrinsic RNA cleavage and catalysis

doi: 10.4161/trns.28869

Figure Lengend Snippet: Figure 3. An intact 3′ - hydroxyl group on the ribose of a 3′-terminal UMP is essential for H1085Y-mediated intrinsic cleavage. ( A ) As outlined, RNAP II EC32 (elongation complex with 32-mer RNA) complexes were divided and their nascent RNAs were extended by 1 nt with either UTP, 3′-deoxy-UTP or 2′-deoxy-UTP. Washes omitting metal cofactors are delineated as “-Me 2+ .” Following a washing step in the absence of metal cofactors, ECs were then exposed to Mn 2+ ions to initiate intrinsic cleavage where appropriate. ( B ) Comparison of cleavage reactions by WT (RNA lengths; 32, 33 nt) and H1085Y (RNA lengths; 32, 33 nt) RNAP II enzymes. A terminal 3′-dUMP prevented RNA hydrolysis by H1085Y RNAP II ( lane 12 ), while the WT RNAP II control lanes reveal little WT RNAP II cleavage activity regardless of the nature of the 3′-NMP added. ( C ) Structures of the tested 3′-NMP modified sugars are depicted. Experiments are representative of at least two independent determinations.

Article Snippet: rNTPs and 2′-deoxy-dATP were obtained from GE Healthcare Life Sciences; [α- 32 P] ATP and [γ- 32 P ] ATP from Perkin-Elmer; 2′-deoxy-UTP, 3′-deoxy-UTP and dinucleotide GpG from TriLink Biotechnologies.

Techniques: Comparison, Control, Activity Assay, Modification