tm3 cells  (Jiancheng Inc)

Journal: Molecular Metabolism

Article Title: AMPK inhibition and elevated angiogenin are associated with tRNA fragmentation in the male germline exposed to a high-fat diet

doi: 10.1016/j.molmet.2026.102350

Figure Lengend Snippet: AMPK inhibition-induced Angiogenin is accompanied by alteration of tRFs profile in vitro. A Western blot results of proteins related with AMPK-mTOR pathways in TM3, TM4, and NIH/3T3 cell lines using compound C (CC), rapamycin (Rapa), and MHY1485 (MHY) in TM3, TM4, and NIH/3T3 cell lines (left to right). The number represents the densiometric comparison of the expression of pAMPK and pmTOR proteins against AMPK and mTOR, respectively. B Angiogenin gene expression. Gene expression was quantified after 12 h of treatment. mRNA expression level was normalized to Actin . Data are represented as mean ± standard deviation. Statistical analysis was performed using One-way ANOVA with Tukey post-hoc test. Different letters are significantly different. C Northern blot results of tRF3-Pro and Val in mouse testis cell lines. Each sample RNAs were extracted from both TM3 and TM4 cells with CC treatment and overexpression of Angiogenin (Ang OE). Each intact tRNA was shown as a control. Band intensities of Northern blot are represented as bar graph after quantification using Image J software. Data are presented as mean ± standard deviation. One-way ANOVA was used for statistical analysis, and the different alphabets with Tukey post-hoc test. D Distribution of read counts based on small RNA type. E Distribution of reads assigned to GtRNAdb in tRF subtypes. The total bar height is proportional to the total read count for each group. Percentages indicate the relative proportion of each tRF subtype within the corresponding group, and n represents the actual read count number for each subtype. tRF5, reads derived from tRNA 5′ end among reads aligned to GtRNAdb; tRF3, reads derived from tRNA 3′ end among reads aligned to GtRNAdb; other, not included in the previous two groups. F Histogram showing the distribution of log 2 (Fold change) using reads aligned to GtRNAdb. Green, CC-treated versus non-treated; Yellow, ANG overexpression versus control. G Volcano plot depicting significantly changed tRFs in TM4 cell lines. Results from CC-treated versus non-treated are represented as circles, and results from ANG overexpression versus control are represented as triangles. n = 2 for each group.

Article Snippet: The mouse testicular TM3 cell line and TM4 Sertoli cell line were purchased from the Korean Cell Line Bank (Seoul, Korea).

Techniques: Inhibition, In Vitro, Western Blot, Comparison, Expressing, Gene Expression, Standard Deviation, Northern Blot, Over Expression, Control, Software, Derivative Assay

Journal: Scientific Reports

Article Title: Morinda officinalis polysaccharides activate the SIRT1/PGC-1α pathway to reduce oxidative damage in Leydig TM3 cells

doi: 10.1038/s41598-026-46267-6

Figure Lengend Snippet: MOP promoted Leydig TM3 cell proliferation. TM3 cells were treated with various concentrations of MOP (50, 100, and 250 mg/L) for 48 h. (A) Cell viability was assessed using the CCK-8 assay. (B) Cell proliferation was evaluated using EdU staining (scale bar = 20 μm). (C) The protein levels of Leydig cell markers (Lhcgr, Insl3, Star, Cyp11a1, Hsd3b1, and Hsd11b1) were determined by Western blot. (D) Testosterone concentration in the culture medium was measured by ELISA. Data are presented as mean ± SD. ** p < 0.05, ** p < 0.01, vs. Control group.

Article Snippet: The levels of superoxide dismutase (SOD; S0101S, Beyotime), catalase (CAT; S0051, Beyotime), glutathione peroxidase (GSH-Px; S0057S, Beyotime), malondialdehyde (MDA; S0131S, Beyotime), and adenosine triphosphate (ATP; A095-1-1, NanJing JianCheng Bioengineering Institute, Jiangsu, China) in various groups of TM3 cells, as well as the levels of testosterone (CSB-E05100r, Wuhan CUSABIO Biotech Co., Ltd, Wuhan, Hubei, China), luteinizing hormone (LH; H206-1-1, JianCheng), and follicle-stimulating hormone (FSH; CSB-E06869r, CUSABIO) in rat serum were detected using the corresponding kits.

Techniques: CCK-8 Assay, Staining, Western Blot, Concentration Assay, Enzyme-linked Immunosorbent Assay, Control

Journal: Scientific Reports

Article Title: Morinda officinalis polysaccharides activate the SIRT1/PGC-1α pathway to reduce oxidative damage in Leydig TM3 cells

doi: 10.1038/s41598-026-46267-6

Figure Lengend Snippet: MOP inhibited H 2 O 2 -caused oxidative damage within Leydig TM3 cells. After Leydig TM3 cells were subjected to 48-h co-treatment with 100 µM H 2 O 2 and MOP, (A) Leydig TM3 cell senescence was detected using SA-β-gal staining; (B) The ROS levels in Leydig TM3 cells were detected using flow cytometry; (C-F) SOD, CAT, GSH-Px, and MDA levels within Leydig TM3 cells were detected using kits. ** p < 0.01, vs. Normal control; ## p < 0.01, vs. the H 2 O 2 group.

Article Snippet: The levels of superoxide dismutase (SOD; S0101S, Beyotime), catalase (CAT; S0051, Beyotime), glutathione peroxidase (GSH-Px; S0057S, Beyotime), malondialdehyde (MDA; S0131S, Beyotime), and adenosine triphosphate (ATP; A095-1-1, NanJing JianCheng Bioengineering Institute, Jiangsu, China) in various groups of TM3 cells, as well as the levels of testosterone (CSB-E05100r, Wuhan CUSABIO Biotech Co., Ltd, Wuhan, Hubei, China), luteinizing hormone (LH; H206-1-1, JianCheng), and follicle-stimulating hormone (FSH; CSB-E06869r, CUSABIO) in rat serum were detected using the corresponding kits.

Techniques: Staining, Flow Cytometry, Control

Journal: Scientific Reports

Article Title: Morinda officinalis polysaccharides activate the SIRT1/PGC-1α pathway to reduce oxidative damage in Leydig TM3 cells

doi: 10.1038/s41598-026-46267-6

Figure Lengend Snippet: MOP inhibited mitochondrial damage in Leydig TM3 cells. (A) Alterations in mitochondrial membrane potential of Leydig TM3 cells were detected using JC-1 dye and flow cytometry; (B) ATP detection kit was used to determine the ATP content in Leydig TM3 cells; (C) Mito-tracker probes (red) were used to stain active mitochondria in Leydig TM3 cells, and a fluorescence inverted microscope was applied to observe the fluorescence intensity; the nucleus was labeled with DAPI (blue). ** p < 0.01, vs. Normal control; # p < 0.05, ## p < 0.01, vs. the H 2 O 2 group.

Article Snippet: The levels of superoxide dismutase (SOD; S0101S, Beyotime), catalase (CAT; S0051, Beyotime), glutathione peroxidase (GSH-Px; S0057S, Beyotime), malondialdehyde (MDA; S0131S, Beyotime), and adenosine triphosphate (ATP; A095-1-1, NanJing JianCheng Bioengineering Institute, Jiangsu, China) in various groups of TM3 cells, as well as the levels of testosterone (CSB-E05100r, Wuhan CUSABIO Biotech Co., Ltd, Wuhan, Hubei, China), luteinizing hormone (LH; H206-1-1, JianCheng), and follicle-stimulating hormone (FSH; CSB-E06869r, CUSABIO) in rat serum were detected using the corresponding kits.

Techniques: Membrane, Flow Cytometry, Staining, Fluorescence, Inverted Microscopy, Labeling, Control

Journal: Scientific Reports

Article Title: Morinda officinalis polysaccharides activate the SIRT1/PGC-1α pathway to reduce oxidative damage in Leydig TM3 cells

doi: 10.1038/s41598-026-46267-6

Figure Lengend Snippet: MOP activated the SIRT1/PGC-1α signaling and upregulated mitochondrial-related functional biomarkers. (A-B) Leydig TM3 cells (with or without H 2 O 2 treatment for 48 h) were treated with 100 mg/L MOP and then detected for the protein levels of SIRT1, PGC-1α, OPA1, TFAM, and BCL2 using Western blot. * p < 0.05, ** p < 0.01, vs. Normal control; ## p < 0.01, vs. the H 2 O 2 group.

Article Snippet: The levels of superoxide dismutase (SOD; S0101S, Beyotime), catalase (CAT; S0051, Beyotime), glutathione peroxidase (GSH-Px; S0057S, Beyotime), malondialdehyde (MDA; S0131S, Beyotime), and adenosine triphosphate (ATP; A095-1-1, NanJing JianCheng Bioengineering Institute, Jiangsu, China) in various groups of TM3 cells, as well as the levels of testosterone (CSB-E05100r, Wuhan CUSABIO Biotech Co., Ltd, Wuhan, Hubei, China), luteinizing hormone (LH; H206-1-1, JianCheng), and follicle-stimulating hormone (FSH; CSB-E06869r, CUSABIO) in rat serum were detected using the corresponding kits.

Techniques: Functional Assay, Western Blot, Control

Journal: Scientific Reports

Article Title: Morinda officinalis polysaccharides activate the SIRT1/PGC-1α pathway to reduce oxidative damage in Leydig TM3 cells

doi: 10.1038/s41598-026-46267-6

Figure Lengend Snippet: Inhibition of SIRT1 reversed the protective effects of MOP against oxidative stress. Leydig TM3 cells were treated with H 2 O 2 (100 µM) alone, or co-treated with MOP (100 mg/L) and the SIRT1 inhibitor EX-527 (10 µM) for 48 h. (A) Cell senescence was evaluated by SA-β-gal staining. (B) Intracellular ROS levels were measured by flow cytometry. (C-F) The levels of SOD, CAT, GSH-Px, and MDA were detected using biochemical kits. (G) Mitochondrial membrane potential was analyzed using JC-1 staining and flow cytometry. (H) ATP content was measured using a biochemical kit. (I) Active mitochondria were visualized using Mito-Tracker Red staining. Data are presented as mean ± SD. ** p < 0.01 vs. H2O2 group; ## p < 0.01 vs. H2O2 + MOP group.

Article Snippet: The levels of superoxide dismutase (SOD; S0101S, Beyotime), catalase (CAT; S0051, Beyotime), glutathione peroxidase (GSH-Px; S0057S, Beyotime), malondialdehyde (MDA; S0131S, Beyotime), and adenosine triphosphate (ATP; A095-1-1, NanJing JianCheng Bioengineering Institute, Jiangsu, China) in various groups of TM3 cells, as well as the levels of testosterone (CSB-E05100r, Wuhan CUSABIO Biotech Co., Ltd, Wuhan, Hubei, China), luteinizing hormone (LH; H206-1-1, JianCheng), and follicle-stimulating hormone (FSH; CSB-E06869r, CUSABIO) in rat serum were detected using the corresponding kits.

Techniques: Inhibition, Staining, Flow Cytometry, Membrane