Journal: Cell Death & Disease

Article Title: CUL4B promotes hepatocellular carcinoma progression and oxaliplatin resistance by facilitating FUS degradation

doi: 10.1038/s41419-025-08320-6

Figure Lengend Snippet: A , B Effect of CUL4B overexpression ( A ) or knockdown ( B ) on the proliferation of Huh7 cells treated with or without 2 μM oxaliplatin. Cell proliferation of HCC cells was determined using a CCK8 growth assay for 4 days (mean ± SD, n = 3). C Effect of CUL4B knockdown on IC 50 of oxaliplatin in Huh7 and LM3 cells. IC 50 was determined using a ATP-lite assay on cells treated with oxaliplatin at indicated concentrations for 48 h (mean ± SD, n = 6). D Effect of CUL4B knockdown on cellular morphology of Huh7 cells treated with 40 μΜ oxaliplatin for 48 h. Cell number was counted (mean ± SD, n = 3). E Effect of CUL4B knockdown on apoptosis of Huh7 cells treated with 40 μΜ oxaliplatin for 48 h. Annexin V-FITC/PI double-staining analysis was used to analyze cell apoptosis (mean ± SD, n = 3). Western blotting was used to assess the expression of the apoptosis-related proteins NOXA and cleaved-PARP. F Effect of CUL4B knockdown on the cell cycle of Huh7 cells treated with 40 μΜ oxaliplatin for 48 h. PI staining and FACS analysis were used to analyze the cell cycle profile (mean ± SD, n = 3). Western blotting was used to assess the expression of the cell cycle-related protein Cyclin B. Two-tailed, unpaired t -test was used for ( A ). One-way ANOVA/LSD test was used for ( B − F ). Data are presented as the mean ± standard deviation derived from a minimum of three independent experiments. Statistical significance was assessed using the appropriate testing methods, with * p < 0.05, ** p < 0.01, and *** p < 0.001 denoting levels of significance.

Article Snippet: For apoptosis assays, the Annexin V-FITC Apoptosis Detection Kit (Beyotime, C1062L) was used according to the manufacturer’s instructions.

Techniques: Over Expression, Knockdown, Growth Assay, Double Staining, Western Blot, Expressing, Staining, Two Tailed Test, Standard Deviation, Derivative Assay

Journal: Oncology Letters

Article Title: Chelerythrine chloride induces apoptosis in renal cancer HEK-293 and SW-839 cell lines

doi: 10.3892/ol.2016.4520

Figure Lengend Snippet: CC-induced dose-dependent apoptosis of HEK-293 and human renal cancer SW-839 cells. (A) HEK-293 and (B) SW-839 cells were treated with various concentrations of CC (0, 5 and 10 µM) for 24 h, and stained with annexin V-fluorescein isothiocyanate/propidium iodide. The percentage of early-stage apoptotic cells is shown in the lower right quadrant, while the percentage of late-stage apoptotic cells is shown in the upper right quadrant. (C) Treatment of HEK-293 and SW-839 cells with 5 and 10 µM CC for 24 h induced cell apoptosis. CC, chelerythrine chloride; FL-H, fluorescence line height.

Article Snippet: One Step TUNEL Apoptosis Assay Kit (Beyotime Institute of Biotechnology, Haimen, China) was used to stain the apoptotic tumor cells.

Techniques: Staining, Fluorescence

Journal: Oncology Letters

Article Title: Chelerythrine chloride induces apoptosis in renal cancer HEK-293 and SW-839 cell lines

doi: 10.3892/ol.2016.4520

Figure Lengend Snippet: Tumor growth suppression in a human renal cancer SW-839 xenograft nude mouse model. (A and B) Tumor volume of CC and control tumors. (C) Tumor weight of CC and control tumors. (D) No significant toxicity to mice was observed following treatment with CC (5 mg/kg/day), according to the body weight of the mice. (E) Representative TUNEL staining (red fluorescence) of SW-839 renal cancer xenografts. (F) Quantification of TUNEL + SW-839 cells in tumor xenografts was performed using cellSens Standard software. The data are presented as the mean ± standard deviation of three experiments. **P<0.01 vs. control cells. (G) IHC staining for Bcl-2 and Bcl-2-associated X protein on tumor sections, and (H) quantification of IHC staining using cellSens Standard software. CC, chelerythrine chloride; TUNEL, terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling; Bcl-2, B-cell lymphoma 2; Bax, Bcl-2-associated X protein; IHC, immunohistochemistry.

Article Snippet: One Step TUNEL Apoptosis Assay Kit (Beyotime Institute of Biotechnology, Haimen, China) was used to stain the apoptotic tumor cells.

Techniques: Control, TUNEL Assay, Staining, Fluorescence, Software, Standard Deviation, Immunohistochemistry, End Labeling

Journal: Oncology Letters

Article Title: Chelerythrine chloride induces apoptosis in renal cancer HEK-293 and SW-839 cell lines

doi: 10.3892/ol.2016.4520

Figure Lengend Snippet: (Aa) Western blot analysis of the expression levels of apoptosis-associated proteins in HEK-293 and human renal cancer SW-839 cells following treatment with CC. Quantification of the expression of various proteins in HEK-293 and SW-839 cells, such as (Ab) p53, (Ac) Bax, (Ad) Bcl-2, (Ae) pro-caspase-3, (Af) cleaved caspase 3 and (Ag) PARP using GAPDH as a control. Multiple bands were observed in the cleaved caspase-3 lane due to non-specific binding of antibodies. (Ab-Ag) Quantification of western blotting (*P<0.05 and **P<0.01 vs. controls). (Ba) Western blot analysis of MAPK and Akt pathways after CC treatment in HEK-293 and SW-839 cells. The quantification of protein expression was performed for (Bb) pERK, (Bc) p-p38, (Bd) p-JNK and (Be) p-AKT, with GADPH as a control. The results are representative of ≥3 independent experiments. Multiple bands were observed in the p-ERK, p-JNK and JNK lanes due to non-specific binding of antibodies. *P<0.05 and **P<0.01 vs. controls. CC, chelerythrine chloride; Bcl-2, B-cell lymphoma 2; Bax, Bcl-2-associated X protein; PARP, poly (adenosine diphosphate-ribose) polymerase; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; p-, phospho-; ERK, extracellular signal-regulated kinase; JNK, c-Jun N-terminal kinase.

Article Snippet: One Step TUNEL Apoptosis Assay Kit (Beyotime Institute of Biotechnology, Haimen, China) was used to stain the apoptotic tumor cells.

Techniques: Western Blot, Expressing, Control, Binding Assay

Journal: Environmental Health Perspectives

Article Title: Prolonged Cadmium Exposure and Osteoclastogenesis: A Mechanistic Mouse and in Vitro Study

doi: 10.1289/EHP13849

Figure Lengend Snippet: The effects of cadmium (Cd) exposure on bone metabolic homeostasis in global Nrf2 knockout ( N r f 2 − / − ) and WT littermate ( N r f 2 + / + ) control mice. (A) Schematic illustration of the experimental design. Male mice (14 wk old) were exposed to cadmium chloride ( CdCl 2 ; 100 mg Cd/L) in drinking water for 16 wk. (B) Representative images of the left femur of mice examined by micro-CT. Scale bar: 0.5 mm . (C) Bone density estimations, including bone volume per tissue volume (BV/TV), trabecular separation (Tb.Sp), trabecular thickness (Tb.Th), trabecular number (Tb.N), total area (Tt.Ar), cortical area (Ct.Ar), structure thickness (St.Th) and cortical area per total area (Ct.Ar/Tt.Ar), were calculated using the Skyscan CT-Analyser software (version 1.1.7; Skyscan CTAn) based on micro-CT measurements. n = 6 – 9 . Values are expressed as mean ± SD . Two-way ANOVA with Bonferroni multiple comparison posttest was used. * p < 0.05 vs. N r f 2 + / + with the same treatment; # p < 0.05 vs. control (Cont) of the same genotype. (D) Results of the three-point bending test performed on the left femurs. The maximum load, breaking load, energy absorption, and stiffness were calculated using Bluehill Elements. n = 4 – 5 . Values are expressed as mean ± SD . Two-way ANOVA with Bonferroni multiple comparison posttest was used for the maximum load and breaking load; Kruskal–Wallis test with Dunn’s multiple comparisons posttest was used for the energy absorption and stiffness. * p < 0.05 vs. N r f 2 + / + with the same treatment. # p < 0.05 vs. Cont of the same genotype. (E) Plasma levels of TRAP5b and CTX-I. n = 5 – 6 . Values are expressed as mean ± SD . Two-way ANOVA with Bonferroni multiple comparison posttest was used. * p < 0.05 vs. N r f 2 + / + with the same treatment; # p < 0.05 vs. Cont of the same genotype. (F) Representative histological images of TRAP staining of the right femur of mice. Black arrow, osteoclasts. Scale bar: 1 mm . (G) Osteoclast number (OcN) and osteoclast surface (OcS) in the femur sections with TRAP staining normalized to bone perimeter (Bpm) and bone surface (BS), respectively. n = 4 – 6 . Values are expressed as mean ± SD . Two-way ANOVA with Bonferroni multiple comparison posttest was used. * p < 0.05 vs. N r f 2 + / + with the same treatment; # p < 0.05 vs. Cont of the same genotype. Summary data are provided in Tables S6–S21. Note: ANOVA, analysis of variance; CTX-I, C-terminal telopeptides of type I; micro-CT, micro-computed tomography; Nrf2, nuclear factor erythroid 2-related factor 2; SD, standard deviation; Trap, tartrate-resistant acid phosphatase; W, weeks; WT, wild type.

Article Snippet: Following centrifugation at 5,000 g at 4°C for 5 min to remove cells, the plasma was collected and stored at − 80 ° C . The activity of TRAP5b was measured using the TRAP Activity Assay Kit (E-BC-K871-M, Elabscience).

Techniques: Knock-Out, Control, Micro-CT, Software, Comparison, Clinical Proteomics, Staining, Standard Deviation

Journal: Environmental Health Perspectives

Article Title: Prolonged Cadmium Exposure and Osteoclastogenesis: A Mechanistic Mouse and in Vitro Study

doi: 10.1289/EHP13849

Figure Lengend Snippet: The impacts of cadmium (Cd) exposure on bone metabolic homeostasis in myeloid-specific Nrf2 knockout [ Nrf2 (M)-KO] and their littermate control (Cont) mice. (A) Schematic illustration of the experimental design. Male mice (14 wk old) were exposed to cadmium chloride ( CdCl 2 ; 100 mg Cd/L) in drinking water for 8 or 16 wk. (B) Representative images of the left femur of mice examined by micro-CT. Scale bar: 0.5 mm . (C) Bone density estimations, including bone volume per tissue volume (BV/TV), trabecular separation (Tb.Sp), trabecular thickness (Tb.Th), trabecular number (Tb.N), total area (Tt.Ar), cortical area (Ct.Ar), structure thickness (St.Th), and cortical area per total area (Ct.Ar/Tt.Ar), were calculated using the Skyscan CT-Analyser software (version 1.1.7; Skyscan CTAn) based on micro-CT measurements. n = 5 – 9 . Values are expressed as mean ± SD . Two-way ANOVA with Bonferroni multiple comparison posttest was used for BV/TV, Tb.Sp, Tb.Th, and Tb.N; Kruskal–Wallis test with Dunn’s multiple comparisons posttest was used for Tt.Ar, Ct.Ar, St.Th, and Ct.Ar/Tt.Ar. * p < 0.05 vs. KI with the same treatment; # p < 0.05 vs. Cont of the same genotype. (D) Representative histological images of TRAP staining of the right femur of mice. Black arrow, osteoclasts. Scale bar: 1 mm . (E) Osteoclast number (OcN) and osteoclast surface (OcS) in the femur sections with TRAP staining normalized to bone perimeter (Bpm) and bone surface (BS), respectively. n = 4 – 5 . Values are expressed as mean ± SD . Two-way ANOVA with Bonferroni multiple comparison posttest was used. * p < 0.05 vs. KI with the same treatment; # p < 0.05 vs. Cont of the same genotype. (F–H) Plasma levels of TRAP5b and CTX-I (F), P1NP and OCN (G), and RANKL and OPG (H). n = 5 – 7 . Values are expressed as mean ± SD . Two-way ANOVA with Bonferroni multiple comparison posttest was used for OcN/Bpm, OcS/BS, TRAP5b, CTX-I, P1NP, and OPG; Kruskal–Wallis test with Dunn’s multiple comparisons posttest was used for OCN and RANKL. * p < 0.05 vs. KI with the same treatment; # p < 0.05 vs. Cont of the same genotype. Summary data are provided in Tables S22–S37. Note: ANOVA, analysis of variance; CTX-I, C-terminal telopeptides of type I; KI, N f e 2 l 2 LoxP / LoxP ; KO, Nrf2 (M)-KO (knockout); micro-CT, micro-computed tomography; Nrf2, nuclear factor erythroid 2-related factor 2; OCN, osteocalcin; OPG, osteoprotegerin; P1NP, procollagen type I N-terminal propeptide; RANKL, receptor activator of nuclear factor kappa-B ligand; SD, standard deviation; Trap, tartrate-resistant acid phosphatase; W, week.

Article Snippet: Following centrifugation at 5,000 g at 4°C for 5 min to remove cells, the plasma was collected and stored at − 80 ° C . The activity of TRAP5b was measured using the TRAP Activity Assay Kit (E-BC-K871-M, Elabscience).

Techniques: Knock-Out, Control, Micro-CT, Software, Comparison, Staining, Clinical Proteomics, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Curcumin enhances the antitumor effect of ABT-737 via activation of the ROS-ASK1-JNK pathway in hepatocellular carcinoma cells

doi: 10.3892/mmr.2015.4715

Figure Lengend Snippet: ROS levels are increased in HepG2 cells following co-treatment with ABT-737 and curcumin. (A) HepG2 cells were separately treated with 1% dimethyl sulfoxide, 2 µ M curcumin, 10 µ M ABT-737 and 2 µ M curcumin+10 µ M ABT-737 for 24 h, following which a 2′,7′-dichlorofluorescin diacetate probe was used to measure the levels of cellular ROS. HepG2 cells were treated with 2 µ M curcumin and 10 µ M ABT-737 for 24 h in the presence or absence of the antioxidant, NAC (10 mM). Subsequently, the levels of HepG2 cell apoptosis were analyzed using (B) Hoechst 33258 staining (white arrows indicate cells undergoing apoptosis with nuclear fragmentation visualized by Hoechst staining), (C) Annexin V-fluorescein isothiocyanate/PI staining and (D) caspase-3 activity detection. Data are expressed as the mean ± standard deviation. * P<0.05. ROS, reactive oxygen species; PI, propidium iodide; NAC, N-acetyl-L-cysteine.

Article Snippet: An Annexin V-Fluorescein Isothiocyanate (FITC)/Propidium Iodide (PI) Apoptosis Detection kit was purchased from Beyotime Institute of Biotechnology (Beijing, China).

Techniques: Staining, Activity Assay, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Curcumin enhances the antitumor effect of ABT-737 via activation of the ROS-ASK1-JNK pathway in hepatocellular carcinoma cells

doi: 10.3892/mmr.2015.4715

Figure Lengend Snippet: Increased levels of ROS promote the sustained activation of JNK, which leads to the apoptosis of HepG2 cells. (A) HepG2 cells were separately treated with 1% dimethyl sulfoxide, 2 µ M curcumin, 10 µ M ABT-737 and 2 µ M curcumin+10 µ M ABT-737 for 24 h, following which the total cellular proteins were extracted. Subsequently, the protein levels of p-JNK1/JNK2 and total JNK1/JNK2 were detected using western blotting. (B) HepG2 cells were treated with 2 µ M curcumin+10 µ M ABT-737 for 24 h in the presence or absence of 10 µ M SP600125 (a JNK inhibitor). Following treatment, apoptosis of the HepG2 cells was assessed using annexin V-fluorescein isothiocyanate/PI staining. (C) HepG2 cells were treated with 2 µ M curcumin+10 µ M ABT-737 for 24 h in the presence or absence of antioxidant, NAC (10 mM). Subsequently, the levels of p-JNK1/JNK2 and total JNK1/JNK2 were detected using western blotting. (D) HepG2 cells were separately transfected with control siRNA or ASK1 siRNA for 24 h, following which the cells were treated with 2 µ M curcumin+10 µ M ABT-737 for 24 h. A Trypan blue exclusion assay was then used to measure the total cell death ratio. * P<0.05 vs. control. ROS, reactive oxygen species; JNK, c-Jun N-terminal kinase; p-, phosphorylated; siRNA, small interfering RNA; PI, propidium iodide.

Article Snippet: An Annexin V-Fluorescein Isothiocyanate (FITC)/Propidium Iodide (PI) Apoptosis Detection kit was purchased from Beyotime Institute of Biotechnology (Beijing, China).

Techniques: Activation Assay, Western Blot, Staining, Transfection, Control, Trypan Blue Exclusion Assay, Small Interfering RNA

Journal: Oncology Letters

Article Title: Hedyotis diffusa willd extract suppresses colorectal cancer growth through multiple cellular pathways

doi: 10.3892/ol.2017.7244

Figure Lengend Snippet: EEHDW inhibits cellular proliferation and induces apoptosis in xenograft tumors. (A) Staining for Ki-67 and TUNEL were performed to examine the in vivo effect of EEHDW on proliferation and apoptosis. The photographs are representative images captured at a magnification of ×400. The percentage of positively stained cells was also quantified. To rule out any non-specific staining, PBS was used to replace the primary antibody as a negative control. Data are expressed as the mean ± standard deviation from 10 individual mice in each group. *P<0.05 vs. control mice. (B) A total of three tumors were randomly selected from each group, and the level of cytochrome C, caspase-3, −9 and PARP in tumor tissues was determined by western blotting to examine the in vivo effect of EEHDW on apoptosis. β-actin was used as the internal control. For each tumor sample, western blotting was performed in triplicate. In densitometric analysis, the expression of the target proteins was normalized to the mean protein expression of control. *P<0.05, vs. control mice. EEHDW, ethanol extract of Hedyotis Diffusa Willd. TUNEL, terminal deoxynucleotidyl-transferase-mediated dUTP nick end labeling; PARP, poly(ADP-ribose)polymerase 1.

Article Snippet: TumorTACS in situ Apoptosis kit (catalog no. 4815–30-K) was purchased from R&D Systems Inc. (Minneapolis, MN, USA).

Techniques: Staining, TUNEL Assay, In Vivo, Negative Control, Standard Deviation, Control, Western Blot, Expressing, End Labeling

Journal: Molecular medicine reports

Article Title: Intraperitoneal injection of thalidomide alleviates early osteoarthritis development by suppressing vascular endothelial growth factor expression in mice.

doi: 10.3892/mmr.2018.8980

Figure Lengend Snippet: Figure 2. mRNA expression levels of VEGF and MMP‑13 in the knee articular cartilage of mice among the Sham, Dmm and Dmm+Th groups (n=4 in each group). (A) Relative mRNA expression levels of VEGF in the medial articular cartilage. (B) Relative mRNA expression levels of MMP‑13 in the medial articular cartilage. The values are presented as the mean ± standard deviation. *P<0.05 compared with the Sham group; #P<0.05 compared with the Dmm group. Dmm, destabilization of the medial meniscus; MMP‑13, matrix metalloproteinase‑13; Th, thalidomide; VEGF, vascular endothelial growth factor.

Article Snippet: An ELISA kit of VEGF (E-EL-M1292c) was purchased from Elabscience Biotechnology Co., Ltd., Wuhan, China.

Techniques: Expressing, Standard Deviation

Journal: Molecular medicine reports

Article Title: Intraperitoneal injection of thalidomide alleviates early osteoarthritis development by suppressing vascular endothelial growth factor expression in mice.

doi: 10.3892/mmr.2018.8980

Figure Lengend Snippet: Figure 3. Immunohistochemical analysis of VEGF expression in the knee articular cartilage of mice among the Sham, Dmm and Dmm+Th groups (n=4 in each group). (A) Immunohistochemistry staining of VEGF in the articular cartilage of the medial tibial plateau (magnification, x400, scale bar=100 µm). (B) Quantification of VEGF positive cells, based on the results of immunohistochemistry staining. The values are presented as the mean ± standard deviation. *P<0.05 compared with the Sham group; #P<0.05 compared with the Dmm group. Dmm, destabilization of the medial meniscus; Th, thalidomide; VEGF, vascular endothelial growth factor.

Article Snippet: An ELISA kit of VEGF (E-EL-M1292c) was purchased from Elabscience Biotechnology Co., Ltd., Wuhan, China.

Techniques: Immunohistochemical staining, Expressing, Immunohistochemistry, Staining, Standard Deviation

Journal: Molecular medicine reports

Article Title: Intraperitoneal injection of thalidomide alleviates early osteoarthritis development by suppressing vascular endothelial growth factor expression in mice.

doi: 10.3892/mmr.2018.8980

Figure Lengend Snippet: Figure 5. ELISA analysis of serum VEGF concentration of mice among the Sham, Dmm and Dmm+Th groups (n=8 in each group). The values are presented as the mean ± standard deviation. *P<0.05 compared with the Sham group; #P<0.05 compared with the Dmm group. Dmm, destabilization of the medial meniscus; Th, thalidomide; VEGF, vascular endothelial growth factor.

Article Snippet: An ELISA kit of VEGF (E-EL-M1292c) was purchased from Elabscience Biotechnology Co., Ltd., Wuhan, China.

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay, Standard Deviation

Journal: Translational Neuroscience

Article Title: The role of melatonin in affecting cognitive dysfunction in acute sleep deprivation mice through the nuclear factor kappaB pathway and oxidative stress

doi: 10.1515/tnsci-2025-0379

Figure Lengend Snippet: Melatonin improved CD in male C57BL/6J ASD mice. MWM test was conducted to detect (a) escape latency, (b) time needed for first crossing, (c) number of crossings through the target platform zone, and (d) time spent in the target platform quadrant within 180 s of mice ( n = 12); (e) open field experiment was performed to assess mouse spontaneous activity ( n = 12); (f) elevated plus maze test was conducted to assess time spent in open and enclosed arms ( n = 12); (g) ELISA was used to measure AChE activity and ACh levels in the hippocampal CA1 region of SD mice ( n = 6). Data in panel (c) failed the Shapiro–Wilk test for normality and the Mann–Whitney U -test was applied for inter-group comparisons. All other data were normally distributed, with the results presented as mean ± standard deviation. (a) Escape latency in the MWM test was analyzed using two-way ANOVA, with the Šídák’s multiple comparison test used for post hoc analyses; (b)–(g) one-way ANOVA was applied for comparisons among multiple groups, and Tukey’s multiple comparison test for post hoc analysis. * P < 0.05, ** P < 0.01.

Article Snippet: ELISA kits were used to determine levels of IL-4 (Mouse IL-4 ELISA Kit, ml063156, Enzyme-linked Biotechnology, Shanghai, China), IL-10 (Mouse IL-10 ELISA Kit, ml037873, Enzyme-linked Biotechnology), TNF-α (Mouse TNF-α ELISA Kit, ml002095, Enzyme-linked Biotechnology), IL-1β (Mouse IL-1β ELISA Kit, ml301814, Enzyme-linked Biotechnology), acetylcholine (ACh) (mouse ACh ELISA kit, E-EL-0081, Elabscience, Wuhan, Hubei.

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Standard Deviation, Comparison

Journal: Translational Neuroscience

Article Title: The role of melatonin in affecting cognitive dysfunction in acute sleep deprivation mice through the nuclear factor kappaB pathway and oxidative stress

doi: 10.1515/tnsci-2025-0379

Figure Lengend Snippet: Melatonin mitigated neuroinflammation and oxidative stress in the hippocampal CA1 region of male C57BL/6J ASD mice. (a) HE staining to observe the structure of the hippocampal CA1 region of mice; (b) Nissl staining to observe the neurons of the hippocampal CA1 region of mice; ELISA to determine TNF-α, IL-1β, IL-4, and IL-10, (c) MDA and ROS levels, and SOD activity in the hippocampal CA1 region of mice (d); (e) western blot to measure PSD95 protein expression in the hippocampal CA1 region of mice; n = 6. All data were normally distributed as determined by the Shapiro–Wilk test and were presented as mean ± standard deviation. One-way ANOVA was applied for comparisons among multiple groups, and Tukey’s multiple comparison for post hoc analysis. Multiple comparisons were subjected to FDR correction using the Benjamini–Hochberg method ( q = 0.05). Corrected P -values were denoted as P adj , while uncorrected raw P -values were labeled as P . FDR correction ( q = 0.05) was applied to all multiple comparisons within each analytical category (inflammatory cytokines, oxidative stress markers). * P < 0.05, ** P < 0.01.

Article Snippet: ELISA kits were used to determine levels of IL-4 (Mouse IL-4 ELISA Kit, ml063156, Enzyme-linked Biotechnology, Shanghai, China), IL-10 (Mouse IL-10 ELISA Kit, ml037873, Enzyme-linked Biotechnology), TNF-α (Mouse TNF-α ELISA Kit, ml002095, Enzyme-linked Biotechnology), IL-1β (Mouse IL-1β ELISA Kit, ml301814, Enzyme-linked Biotechnology), acetylcholine (ACh) (mouse ACh ELISA kit, E-EL-0081, Elabscience, Wuhan, Hubei.

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Activity Assay, Western Blot, Expressing, Standard Deviation, Comparison, Labeling

Journal: Translational Neuroscience

Article Title: The role of melatonin in affecting cognitive dysfunction in acute sleep deprivation mice through the nuclear factor kappaB pathway and oxidative stress

doi: 10.1515/tnsci-2025-0379

Figure Lengend Snippet: Suppression of the NF-κB pathway improved neuroinflammation and oxidative stress in the hippocampal CA1 region of male C57BL/6J ASD mice. (a/f) Western blot to determine p-p65/p65 ratio and PSD95 protein expression in the hippocampal CA1 region of mice; (b) HE staining to observe the structure of the hippocampal CA1 region of mice; (c) Nissl staining to observe the neuronal status of the hippocampal CA1 region of mice; ELISA to determine (d) TNF-α, IL-1β, IL-4, and IL-10 levels and (e) MDA levels, ROS, and SOD activity in the hippocampal CA1 region; n = 6. Data were expressed as mean ± standard deviation. The t -test was adopted for inter-group comparisons. Multiple comparisons were subjected to FDR correction using the Benjamini–Hochberg method ( q = 0.05). Corrected P -values were denoted as P adj , while uncorrected raw P -values were labeled as P . FDR correction ( q = 0.05) was applied to all multiple comparisons within each analytical category (inflammatory cytokines, oxidative stress markers). * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: ELISA kits were used to determine levels of IL-4 (Mouse IL-4 ELISA Kit, ml063156, Enzyme-linked Biotechnology, Shanghai, China), IL-10 (Mouse IL-10 ELISA Kit, ml037873, Enzyme-linked Biotechnology), TNF-α (Mouse TNF-α ELISA Kit, ml002095, Enzyme-linked Biotechnology), IL-1β (Mouse IL-1β ELISA Kit, ml301814, Enzyme-linked Biotechnology), acetylcholine (ACh) (mouse ACh ELISA kit, E-EL-0081, Elabscience, Wuhan, Hubei.

Techniques: Western Blot, Expressing, Staining, Enzyme-linked Immunosorbent Assay, Activity Assay, Standard Deviation, Labeling

Journal: Translational Neuroscience

Article Title: The role of melatonin in affecting cognitive dysfunction in acute sleep deprivation mice through the nuclear factor kappaB pathway and oxidative stress

doi: 10.1515/tnsci-2025-0379

Figure Lengend Snippet: Inhibition of the NF-κB pathway notably attenuated CD in male C57BL/6J ASD mice. MWM test to determine (a) escape latency, (b) first crossing time, (c) number of crossings through the target platform zone, and (d) time spent in the target platform quadrant within 180 s of mice ( n = 12); (e) open field experiment to assess spontaneous activity of mice ( n = 12); (f) elevated plus maze test was conducted to assess time spent in open and enclosed arms ( n = 12); (g) ELISA was used to measure AChE activity and ACh levels in the hippocampal CA1 region of SD mice ( n = 6). Data were expressed as mean ± standard deviation. (a) Escape latency in the MWM test was analyzed using two-way ANOVA, with the Šídák’s multiple comparison test used for post hoc analyses; (b)–(g) the t test was utilized for inter-group comparisons. ** P < 0.01.

Article Snippet: ELISA kits were used to determine levels of IL-4 (Mouse IL-4 ELISA Kit, ml063156, Enzyme-linked Biotechnology, Shanghai, China), IL-10 (Mouse IL-10 ELISA Kit, ml037873, Enzyme-linked Biotechnology), TNF-α (Mouse TNF-α ELISA Kit, ml002095, Enzyme-linked Biotechnology), IL-1β (Mouse IL-1β ELISA Kit, ml301814, Enzyme-linked Biotechnology), acetylcholine (ACh) (mouse ACh ELISA kit, E-EL-0081, Elabscience, Wuhan, Hubei.

Techniques: Inhibition, Activity Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation, Comparison

Journal: Translational Neuroscience

Article Title: The role of melatonin in affecting cognitive dysfunction in acute sleep deprivation mice through the nuclear factor kappaB pathway and oxidative stress

doi: 10.1515/tnsci-2025-0379

Figure Lengend Snippet: Activation of the NF-κB signaling promoted oxidative stress and CD in the hippocampal CA1 region of male C57BL/6J ASD mice. (a/f) Western blot for determination of p-p65, p65, and PSD95 protein expression levels in the hippocampal CA1 region ( n = 6); (b) HE staining for observation of the structure of the mouse hippocampal CA1 region ( n = 6); (c) Nissl staining for assessment of neuronal status in the mouse hippocampal CA1 region ( n = 6); (d) ELISA for measurement of levels of TNF-α, IL-1β, IL-4, and IL-10 levels in the hippocampal CA1 region of mice ( n = 6); (e) levels of MDA, ROS, and SOD activity in the mouse hippocampal CA1 region ( n = 6); (f) western blot for determination of PSD95 protein expression; MWM test to detect (g) escape latency, (h) time needed for first crossing, (i) number of crossings through the target platform zone within 180 s, and (j) time spent in the target platform quadrant within 180 s of mice ( n = 12); (k) open field experiment to assess mouse spontaneous activity ( n = 12); (l) elevated plus maze test to quantify time spent in open and enclosed arms ( n = 12); (m) ELISA was used to measure AChE activity and ACh levels in the hippocampal CA1 region of SD mice ( n = 6). Data were presented as mean ± standard deviation. (g) Escape latency was analyzed using two-way ANOVA, with the Šídák’s multiple comparison test used for post hoc analyses; other panels: inter-group comparisons were analyzed by the t -test. Multiple comparisons underwent FDR correction via the Benjamini–Hochberg method ( q = 0.05). Corrected P -values were denoted as P adj and uncorrected raw P -values as P . FDR correction ( q = 0.05) was applied to all multiple comparisons within each analytical category (inflammatory cytokines, oxidative stress markers). * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: ELISA kits were used to determine levels of IL-4 (Mouse IL-4 ELISA Kit, ml063156, Enzyme-linked Biotechnology, Shanghai, China), IL-10 (Mouse IL-10 ELISA Kit, ml037873, Enzyme-linked Biotechnology), TNF-α (Mouse TNF-α ELISA Kit, ml002095, Enzyme-linked Biotechnology), IL-1β (Mouse IL-1β ELISA Kit, ml301814, Enzyme-linked Biotechnology), acetylcholine (ACh) (mouse ACh ELISA kit, E-EL-0081, Elabscience, Wuhan, Hubei.

Techniques: Activation Assay, Western Blot, Expressing, Staining, Enzyme-linked Immunosorbent Assay, Activity Assay, Standard Deviation, Comparison

Journal: Endocrinology and Metabolism

Article Title: Fancd2os Reduces Testosterone Production by Inhibiting Steroidogenic Enzymes and Promoting Cellular Apoptosis in Murine Testicular Leydig Cells

doi: 10.3803/enm.2022.1431

Figure Lengend Snippet: Fig. 4. Fancd2 opposite-strand (Fancd2os) reduces testosterone production and steroidogenic enzyme expression in TM3 cells. The testoster one levels in Fancd2os-overexpressing (A) or knockdown TM3 cells (B) were detected by enzyme-linked immunosorbent assays (n=3 per group). (C, D, E) Relative quantities of mRNA expression of steroidogenic acute regulatory protein (StAR), P450 cholesterol side-chain cleavage (P450scc), and 3β-hydroxysteroid dehydrogenase (3β-HSD) in Fancd2os-overexpressing TM3 cells or Fancd2os knockdown TM3 cells (F, G, H) were determined real-time polymerase chain reaction using β-actin as a housekeeping gene. Each bar represents the mean± standard deviation from three separate experiments. Significant difference compared to TM3, vector/TM3 or NC/TM3. aP<0.05; bP<0.01.

Article Snippet: The testosterone concentration of both serum and cell supernatants was measured using a testosterone ELISA Kit (Elabscience, Houston, TX, USA) according to the manufacturer’s protocol.

Techniques: Expressing, Knockdown, Real-time Polymerase Chain Reaction, Standard Deviation, Plasmid Preparation

Journal: Endocrinology and Metabolism

Article Title: Fancd2os Reduces Testosterone Production by Inhibiting Steroidogenic Enzymes and Promoting Cellular Apoptosis in Murine Testicular Leydig Cells

doi: 10.3803/enm.2022.1431

Figure Lengend Snippet: Fig. 6. Higher Fancd2 opposite-strand (Fancd2os) levels in older mouse Leydig cells result in cellular apoptosis and lower serum testosterone production. (A) The serum testosterone levels from mice of different ages were measured using enzyme-linked immunosorbent assays. (B, C) The testis tissues from different aged mice were sliced and then Fancd2os protein expression and apoptosis were analyzed using immuno chemistry and the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay, respectively. ST represents seminiferous tubule. Black arrows and red arrows indicate Fancd2os-positive cells and TUNEL-positive cells, respectively. Scale bar repre sents 25 μm. The results are given as the mean±standard deviation (n=3). (D) The correlations between Fancd2os expression (B) and the TUNEL-positive staining rate (C) were analyzed using Pearson correlation coefficients. An r≥0.5 was considered to indicate a strong correla tion, and P<0.05 was considered statistically significant. aP<0.05 compared with juvenile mice; bP<0.05 compared with young mice; cP<0.05 compared with middle-aged mice.

Article Snippet: The testosterone concentration of both serum and cell supernatants was measured using a testosterone ELISA Kit (Elabscience, Houston, TX, USA) according to the manufacturer’s protocol.

Techniques: Expressing, End Labeling, TUNEL Assay, Standard Deviation, Staining

Journal: Molecular Medicine Reports

Article Title: Continuous expression of CD83 on activated human CD4 + T cells is correlated with their differentiation into induced regulatory T cells

doi: 10.3892/mmr.2015.3796

Figure Lengend Snippet: Time-dependent expression of CD83, proliferation and the production of IL-2 and IFN-γ in CD4 + T cells. Following purity identification, the purified CD4 + T cells were either (A) directly stained with PE-labeled CD83 or (B–D) stimulated with anti-CD3/CD28 for 1–3 days, followed by staining with PE-labeled CD83. All samples were analyzed by flow cytometry and the typical flow cytometric histograms indicating the percentages of CD83-positive cells are shown. (E) Additionally, cell proliferation at days 1, 2 and 3 were determined using Cell Counting Kit 8. (F) The levels of IL-2 and IFN-γ in the supernatants from 1-, 2- and 3-day cultures of activated CD4 + T cells were analyzed by ELISA. Values are expressed as the mean ± standard deviation of three independent experiments ( ** P<0.01; * P<0.05). PE, phycoerythrin; CD, cluster of differentiation; IFN, interferon; IL, interleukin; neg.con, negative control.

Article Snippet: The levels of IL-2 and IFN-γ in CD4 + T-cell culture supernatants were measured in duplicate for each of the serial aliquots using a Human IL-2 Quantikine ELISA kit (cat. no. D2050) and Human IFN-γ Quantikine ELISA kit (cat. no. DIF50), purchased from R&D Systems (Minneapolis, MN, USA), according to the manufacturer's instructions.

Techniques: Expressing, Purification, Staining, Labeling, Flow Cytometry, Cell Counting, Enzyme-linked Immunosorbent Assay, Standard Deviation, Negative Control