mouse sertoli cell line tm4 gnm41  (Procell Inc)

Journal: Toxics

Article Title: PFOA Damages Blood–Testis Barrier Integrity in Mice by Inhibited Glycolysis Caused H3K18 Lactylation Modification Impairment

doi: 10.3390/toxics14050399

Figure Lengend Snippet: PFOA altered the expression of glycolysis-related genes. Based on RNA-seq analysis, the total RNA expression profiles of testicular tissues in the control and 5 mg/kg/day PFOA-exposed groups were compared ( A ). KEGG pathway enrichment ( B ) and GO functional enrichment analyses ( C ) of differentially expressed genes revealed that the glycolysis pathway served as a key target of PFOA. After 24 h of PFOA treatment in TM4 cells, cell viability was decreased ( D ), accompanied by significant changes in the expression of glycolysis-related genes ( E ). ( n = 3, * p < 0.05; ** p < 0.01; *** p < 0.001, * vs. control group).

Article Snippet: This experiment utilized a mouse testicular sertoli cell line (TM4) obtained from the ATCC cell bank in the United States.

Techniques: Expressing, RNA Sequencing, RNA Expression, Control, Functional Assay

Journal: Toxics

Article Title: PFOA Damages Blood–Testis Barrier Integrity in Mice by Inhibited Glycolysis Caused H3K18 Lactylation Modification Impairment

doi: 10.3390/toxics14050399

Figure Lengend Snippet: PFOA inhibited the expression of glycolysis and BTB-related molecules in TM4 cells and mouse testicular tissues. In PFOA-treated TM4 cells, the protein levels of key glycolytic molecules, including HK1 ( B ), GLUT1 ( C ) and LDHA ( D ), were downregulated ( A – D ), accompanied by reduced production of lactate ( E ) and ATP ( F ). Meanwhile, both the gene ( G ) and protein ( H ) expression of BTB-related factors were decreased in TM4 cells. Consistent with the cellular results, the mRNA ( I ) and protein ( J ) levels of glycolysis-related molecules were also suppressed in mouse testis, along with a decline in lactate concentration ( K ). ( n = 3, * p < 0.05; ** p < 0.01; *** p < 0.001, * vs. control group).

Article Snippet: This experiment utilized a mouse testicular sertoli cell line (TM4) obtained from the ATCC cell bank in the United States.

Techniques: Expressing, Concentration Assay, Control

Journal: Toxics

Article Title: PFOA Damages Blood–Testis Barrier Integrity in Mice by Inhibited Glycolysis Caused H3K18 Lactylation Modification Impairment

doi: 10.3390/toxics14050399

Figure Lengend Snippet: Effects of PFOA on H3K18 lactylation and the transcriptional regulation of key genes involved in glycolysis. PFOA exposure reduced the modification levels of Pan-Kla and H3K18la in TM4 cells ( A ) and mouse testicular tissues ( B ). ChIP-qPCR results further demonstrated that PFOA altered the binding of H3K18la to glycolytic genes, thereby regulating their transcription ( C ). ( n = 3, * p < 0.05; ** p < 0.01; *** p < 0.001, * vs. control group).

Article Snippet: This experiment utilized a mouse testicular sertoli cell line (TM4) obtained from the ATCC cell bank in the United States.

Techniques: Modification, ChIP-qPCR, Binding Assay, Control

Journal: Toxics

Article Title: PFOA Damages Blood–Testis Barrier Integrity in Mice by Inhibited Glycolysis Caused H3K18 Lactylation Modification Impairment

doi: 10.3390/toxics14050399

Figure Lengend Snippet: Protective effects of Nala against PFOA-induced reproductive damage. Gradient Nala treatment for 24 h significantly elevated the levels of Pan-Kla and H3K18la in TM4 cells ( A ). After co-treatment with 30 mM Nala and 400 μM PFOA for 24 h, the PFOA-mediated reductions in Pan-Kla and H3K18la were notably restored ( B ). The decreased expression of Hk1, Glut1 and Ldha was partially rescued ( C ), and the downregulated mRNA ( D ) and protein ( E ) levels of BTB-related molecules were also markedly recovered. ( n = 3, * p < 0.05; ** p < 0.01; *** p < 0.001; * vs. control group. # p < 0.05; ## p < 0.01; ### p < 0.001; # vs. PFOA group).

Article Snippet: This experiment utilized a mouse testicular sertoli cell line (TM4) obtained from the ATCC cell bank in the United States.

Techniques: Expressing, Control

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: Research

Article Title: Environmental Fluoride Compromises Male Fertility: Differentially Modulated miR-34a-5p Targets REST to Regulate Autophagy in Testicular Somatic Cells

doi: 10.34133/research.1113

Figure Lengend Snippet: Fluoride impairs male reproductive capacity by differentially regulating autophagy of testicular somatic cells in mice. (A to F) Six-week-old male mice freely drank distilled water with or without 100 mg/l NaF for 18 weeks. (A) Representative images of H&E staining on testis. Scale bars, 100 μm (left) and 50 μm (right). (B) Representative microscopic views of sperm morphology. Scale bars, 50 μm. (C) Sperm quality. (D and E) Representative transmission electron microscopy images of autophagosomes in (D) Leydig cells and (E) Sertoli cells [scale bars, 2 μm (bottom) and 1 μm (top)], with red arrows indicating autophagosomes. (F) Immunohistochemistry was used to detect the protein expression of LC3B and p62 in Leydig cells and Sertoli cells. (G and H) Testicular somatic cells were treated with various concentrations of NaF for 24 h. The relative expression of LC3B and p62 proteins in (G) TM3 cells, (H) TM4 cells, and (I) TM4 cells cotreated with 20 μM chloroquine. Cell viability of (J) TM3 and (K) TM4 cells that were treated with 0.25 mM NaF and 20 μM chloroquine for 24 h or pretreated with 20 nM rapamycin for 1 h. All values in the figure are means ± SEM. n ≥ 3. * P < 0.05, ** P < 0.01 versus control; # P < 0.05, ## P < 0.01 versus chloroquine.

Article Snippet: Murine Leydig cell line (TM3) and murine Sertoli cell line (TM4) were purchased from the American Type Culture Collection (ATCC) (Manassas, USA).

Techniques: Staining, Transmission Assay, Electron Microscopy, Immunohistochemistry, Expressing, Control

Journal: Research

Article Title: Environmental Fluoride Compromises Male Fertility: Differentially Modulated miR-34a-5p Targets REST to Regulate Autophagy in Testicular Somatic Cells

doi: 10.34133/research.1113

Figure Lengend Snippet: miR-34a-5p was selected as the key miRNA in the differential regulation of autophagy in testicular somatic cells by fluoride. (A) Network toxicological analysis results of miRNAs related to fluoride-induced male reproductive injury. (B) HAMDB database analysis results of autophagy-related candidate miRNAs. (C and D) qRT-PCR revealed autophagy-related candidate miRNA expression changes in (C) TM3 and (D) TM4 cells after fluoride treatment. (E) FISH of miR-34a-5p in the testis of control and fluoride-treated mice (dashed areas indicate Leydig cells, and white arrows indicate Sertoli cells). All values in the figure are means ± SEM. n = 3. * P < 0.05, ** P < 0.01.

Article Snippet: Murine Leydig cell line (TM3) and murine Sertoli cell line (TM4) were purchased from the American Type Culture Collection (ATCC) (Manassas, USA).

Techniques: Quantitative RT-PCR, Expressing, Control

Journal: Research

Article Title: Environmental Fluoride Compromises Male Fertility: Differentially Modulated miR-34a-5p Targets REST to Regulate Autophagy in Testicular Somatic Cells

doi: 10.34133/research.1113

Figure Lengend Snippet: Fluoride differentially regulates miR-34a-5p to modulate autophagy in testicular somatic cells. (A to F) Testicular somatic cells were treated with lentiviruses expressing miR-34a-5p/miR-NC and Inhibitor-miR-34a-5p/Inhibitor-NC. (A and D) Relative expression of miR-34a-5p. (B, C, E, and F) Relative expression of LC3B and p62 proteins. (G to I) miR-34a-5p overexpression rescued fluoride-treated TM3 cells. (G) Relative expression of miR-34a-5p. (H and I) Relative expression of LC3B and p62 proteins. (J to L) miR-34a-5p knockdown rescued fluoride-treated TM4 cells. (J) Relative expression of miR-34a-5p. (K and L) Relative expression of LC3B and p62 proteins. All values in the figure are means ± SEM. n = 3. * P < 0.05, ** P < 0.01 versus control, miR-NC, or Inhibitor-NC; # P < 0.05, ## P < 0.01 versus 0.25 mM NaF.

Article Snippet: Murine Leydig cell line (TM3) and murine Sertoli cell line (TM4) were purchased from the American Type Culture Collection (ATCC) (Manassas, USA).

Techniques: Expressing, Over Expression, Knockdown, Control

Journal: Research

Article Title: Environmental Fluoride Compromises Male Fertility: Differentially Modulated miR-34a-5p Targets REST to Regulate Autophagy in Testicular Somatic Cells

doi: 10.34133/research.1113

Figure Lengend Snippet: miR-34a-5p targets REST in testicular somatic cells exposed to fluoride. (A) Expression changes of p62 mRNA in TM3 and TM4 cells after fluoride treatment and overexpression or knockdown of miR-34a-5p. (B) Prediction results of miR-34a-5p target genes by miRWalk combined with ChEA3 database. (C and F) Differential expression of the top 15 predicted target genes after fluoride treatment in (C) TM3 and (F) TM4 cells. (D, E, G, and H) Relative mRNA expression of Elf1 , Foxo1 , Foxp1 , Klf10 , Mef2a , Rest , Smad5 , and Yy1 . (I) Schematic showing the targeted binding sites of miR-34a-5p to FOXO1 and REST 3′ UTR. (J and K) The histogram shows the relative fluorescence intensity of each group. (L) FISH of miR-34a-5p and Rest mRNA in testicular tissue from control mice (white arrow indicates the positive area within the Sertoli cells or Leydig cells). (M and N) Differential expression of REST protein in testicular somatic cells after overexpression or knockdown of miR-34a-5p. All values in the figure are means ± SEM. n = 3. * P < 0.05, ** P < 0.01.

Article Snippet: Murine Leydig cell line (TM3) and murine Sertoli cell line (TM4) were purchased from the American Type Culture Collection (ATCC) (Manassas, USA).

Techniques: Expressing, Over Expression, Knockdown, Quantitative Proteomics, Binding Assay, Fluorescence, Control

Journal: Research

Article Title: Environmental Fluoride Compromises Male Fertility: Differentially Modulated miR-34a-5p Targets REST to Regulate Autophagy in Testicular Somatic Cells

doi: 10.34133/research.1113

Figure Lengend Snippet: miR-34a-5p targets REST to regulate autophagy in testicular somatic cells. (A to F) TM3 and TM4 cells were treated with lentivirus expressing OE-REST, si-REST, OE-NC, or si-NC. (A and D) Relative Rest mRNA expression. (B, C, E, and F) Relative LC3B, p62, and REST protein expression. (G to K) REST overexpression rescued TM3 and TM4 cells overexpressing miR-34a-5p. (G and H) Relative miR-34a-5p, Rest , and p62 mRNA expression. (I to K) Relative REST, LC3B, and p62 protein levels. All values in the figure are means ± SEM. n = 3. ns indicates not significant; * P < 0.05, ** P < 0.01 versus control; # P < 0.05, ## P < 0.01 versus miR-34a-5p.

Article Snippet: Murine Leydig cell line (TM3) and murine Sertoli cell line (TM4) were purchased from the American Type Culture Collection (ATCC) (Manassas, USA).

Techniques: Expressing, Over Expression, Control

Journal: Research

Article Title: Environmental Fluoride Compromises Male Fertility: Differentially Modulated miR-34a-5p Targets REST to Regulate Autophagy in Testicular Somatic Cells

doi: 10.34133/research.1113

Figure Lengend Snippet: Fluoride differentially regulates REST to modulate autophagy in testicular somatic cells. (A) Representative image of Rest mRNA FISH in testes from control and fluoride-treated mice. (B) Representative immunofluorescence staining of REST proteins in testes from control and fluoride-treated mice. Dashed areas indicate Leydig cells, and white arrows indicate Sertoli cells. (C and D) Relative REST protein expression in testicular somatic cells treated with fluoride. (E to G) REST knockdown rescued fluoride-treated TM3 cells. (E) Relative expression of Rest mRNA. (F and G) Relative expression of REST, p62, and LC3B proteins. (H to J) REST overexpression rescued fluoride-treated TM4 cells. (H) Relative expression of Rest mRNA. (I and J) Relative expression of REST, p62, and LC3B proteins. All values in the figure are means ± SEM. n = 3. * P < 0.05, ** P < 0.01 versus control, si-NC, or OE-NC; # P < 0.05, ## P < 0.01 versus 0.25 mM NaF.

Article Snippet: Murine Leydig cell line (TM3) and murine Sertoli cell line (TM4) were purchased from the American Type Culture Collection (ATCC) (Manassas, USA).

Techniques: Control, Immunofluorescence, Staining, Expressing, Knockdown, Over Expression

Journal: Cell Biology and Toxicology

Article Title: ATG5-dependent autophagy in Sertoli cells protects against cadmium-disrupted blood-testis barrier via perturbing CXCL2/CXCR2 axis

doi: 10.1007/s10565-025-10089-5

Figure Lengend Snippet: Cd exposure activates autophagy in Sertoli cells. A - G The male mice were injected (i.p.) with 2.0 mg/kg CdCl 2 or vehicle, respectively. Mouse testes were collected at 0, 2, 8, 24, 48, and 72 h after Cd treatment. A , B Representative images and quantification of autophagosomes in mice Sertoli cells detected by transmission electron microscopy. The yellow arrows represent the autophagosomes. C - E The expression of ATG5 and LC3B-II/I was detected by western blotting. F , G Representative images and quantification of mIHC assay. White arrows denoted the LC3B puncta surrounding WT1. Scale bar: 50 μm. H - J TM4 cells were exposed to Cd for 0, 2, 8, and 24 h, respectively. H - J The expression of ATG5 and LC3B-II/I was detected by western blotting. Data were displayed as mean ± SEM . n = 3- 6. * P < 0.05, ** P < 0.01 vs 0 h group

Article Snippet: The mouse Sertoli cell line TM4 (GNM41) was obtained from Procell Life Science & Technology(Li et al. ).

Techniques: Injection, Transmission Assay, Electron Microscopy, Expressing, Western Blot