Journal: Nutrients

Article Title: Functionally Relevant Differences in Plasma Fatty Acid Composition and Expression of Cytotoxic and Inhibitory NK Cell Receptors between Healthy Young and Healthy Elder Adults

doi: 10.3390/nu12123641

Figure Lengend Snippet: Expression of Killing Inhibitory Receptors (KIR) and activating receptors in NK cells derived from young adults and elders. The results represent the percentage mean and SD and Mean Channel Fluorescence Intensity (MFI) in linear units for each group ( n = 30). The statistical analysis was accomplished using the paired Student’s t -test in both parameters analysed.

Article Snippet: The antibodies anti-CD158e1/e2 KIR3DL1 (clone REA168), anti-KIR2DS4/CD158f (clone JJC11.6) were purchased from Miltenyi Biotech.

Techniques: Expressing, Derivative Assay, Fluorescence

Journal: Frontiers in Cellular Neuroscience

Article Title: Adipose-derived stem cell-conditioned medium mitigates ischemia-induced neuronal injury via the JAK1/STAT3 signaling pathway

doi: 10.3389/fncel.2026.1744887

Figure Lengend Snippet: ADSC-CM promotes neurite outgrowth in OGD-injured neurons through the JAK1-STAT3 signaling pathway. (A) On day 5 of cultivation, neuronal neurites extended and formed an extensive neural network. Immunofluorescence staining showed that the neurons were positive for the neuronal-specific marker Tuj1. After OGD injury, neuronal neurites were damaged, partially interrupted, and disappeared. (B) Western blot bands of JAK1, pJAK1, STAT3, pSTAT3, and GAPDH for each group. (C) Comparison of the relative ratio of pJAK1 to JAK1 across experimental groups ( n = 4). (D) Comparison of the relative ratio of pSTAT3 to STAT3 across experimental groups ( n = 4). (E) Immunofluorescence images of neurons from each group, showing the effects of ADSC-CM and GLPG0634 on neurite outgrowth. Neurons were stained with the Tuj1 antibody (red) to label the neuronal cytoskeleton, and nuclei were counterstained with DAPI (blue). (F) Diagram illustrating how to measure the length of the longest neurite and the number of primary neurites in neurons. The green line shows the trajectory of the longest neurite, and white arrows point to primary neurites. (G,H) Comparison of the length of the longest neurite and the number of primary neurites in neurons across experimental groups ( n = 4). Data are expressed as means ± SEM. The difference between the groups was assessed using a one-way ANOVA followed by Bonferroni post hoc tests. * p < 0.05, ** p < 0.01, *** p < 0.001. Scale bars: 100 μm (A) , 20 μm (E,F) .

Article Snippet: After overnight incubation with rabbit anti-Tuj1 antibody (1:200, BM3881, Boster, Wuhan, China) at 4 °C, cells were washed and then incubated with Cy3-conjugated goat anti-rabbit IgG (1:400, AS007, ABclonal) for 1 h at room temperature in the dark.

Techniques: Immunofluorescence, Staining, Marker, Western Blot, Comparison

Journal: Oncology Research

Article Title: BHLHE40 Is a Transcriptional Regulatory Target of NFE2L3 in Triple-Negative Breast Cancer

doi: 10.32604/or.2025.070793

Figure Lengend Snippet: ( A ) Consensus regulatory targets of NFE2L3, based on the regulatory networks inferred using GENIE3 and ARACNe. The red represents the protein upregulated in TNBC compared to TPBC. The blue represents the protein downregulated in TNBC compared to TPBC. Kaplan-Meier survival plots of (B) NFE2L3 and (C) BHLHE40 using the TCGA RNA-seq Breast cancer dataset. The red line represents higher expression of the gene, while the blue line represents lower expression of the gene. p < 0.05. Higher NFE2L3 expression corresponds to poor overall survival than those with low NFE2L3 expression, while there is no significant difference in survival between patients with high and low expressions of BHLHE40

Article Snippet: The membranes were washed with PBST solution for 5 min three times, and then incubated with primary antibodies against NFE2L3 (rabbit polyclonal antibody; #A09888, Boster Bio, Pleasanton, CA, USA), BHLHE40 (rabbit polyclonal antibody; #ABN1737, Millipore-Sigma, Burlington, MA), and GAPDH (rabbit monoclonal antibody; #2118, Cell Signaling Technology, Danvers, MA, USA), were diluted in blocking solution at 4°C overnight.

Techniques: RNA Sequencing, Expressing

Journal: Oncology Research

Article Title: BHLHE40 Is a Transcriptional Regulatory Target of NFE2L3 in Triple-Negative Breast Cancer

doi: 10.32604/or.2025.070793

Figure Lengend Snippet: Gene expression quantification using quantitative Reverse Transcriptase polymerase chain reaction (qRT-PCR). NFE2L3 gene expression in ( A ) MDA-MB-231 and ( B ) MDA-MB-468 cells in siControl, siNFE2L3, and siBHLHE40 groups. ( C , D ) Gene expression in ( C ) MDA-MB-231 and ( D ) MDA-MB-468 cells in untreated, NFE2L3 plasmid and BHLHE40 plasmid groups. BHLHE40 gene expression using qRT-PCR. BHLHE40 gene expression in ( E ) MDA-MB-231 and ( F ) MDA-MB-468 cells following treatment with siControl, siNFE2L3, and siBHLHE40. BHLHE40 gene expression in ( G ) MDA-MB-231 and ( H ) MDA-MB-468 cells following treatment with vehicle (untreated), NFE2L3 plasmid, and BHLHE40 plasmid. One-way ANOVA followed by post-hoc Tukey’s test was used. Compared to the siControl or untreated group: ***indicates p < 0.001, and ****indicates p < 0.0001. Compared to siNFE2L3 or NFE2L3 plasmid group: #### indicates p < 0.0001

Article Snippet: The membranes were washed with PBST solution for 5 min three times, and then incubated with primary antibodies against NFE2L3 (rabbit polyclonal antibody; #A09888, Boster Bio, Pleasanton, CA, USA), BHLHE40 (rabbit polyclonal antibody; #ABN1737, Millipore-Sigma, Burlington, MA), and GAPDH (rabbit monoclonal antibody; #2118, Cell Signaling Technology, Danvers, MA, USA), were diluted in blocking solution at 4°C overnight.

Techniques: Gene Expression, Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Plasmid Preparation

Journal: Oncology Research

Article Title: BHLHE40 Is a Transcriptional Regulatory Target of NFE2L3 in Triple-Negative Breast Cancer

doi: 10.32604/or.2025.070793

Figure Lengend Snippet: Basal protein expression of NFE2L3 and BHLHE40 NFE2L3 and BHLHE40 in breast cancer cells. ( A ) Representative blots of NFE2L3 and BHLHE40 protein expression following treatment with siControl, siNFE2L3, and siBHLHE40. GAPDH was used as a loading control. ( B ) Graphical presentation of NFE2L3 protein expression in MDA-MB-231 cells following treatment as described. ( C ) Graphical presentation of BHLHE40 protein expression in MDA-MB-231 cells following treatment as described. ( D ) Representative blots of NFE2L3 and BHLHE40 protein expression following treatment with vehicle (Untreated), NFE2L3 DNA plasmid, and BHLHE40 DNA plasmid. GAPDH was used as a loading control. ( E ) Graphical presentation of NFE2L3 protein expression in MDA-MB-231 cells following treatment as described. ( F ) Graphical presentation of BHLHE40 protein expression in MDA-MB-231 cells following treatment as described. One-way ANOVA followed by post-hoc Dunnet’s test was used. Compared to siControl: **indicates p < 0.01, ***indicates p < 0.001, and ****indicates p < 0.0001. Compared to the siNFE2L3 group or NFE2L3 plasmid group: # indicates p < 0.05, and #### indicates p < 0.0001

Article Snippet: The membranes were washed with PBST solution for 5 min three times, and then incubated with primary antibodies against NFE2L3 (rabbit polyclonal antibody; #A09888, Boster Bio, Pleasanton, CA, USA), BHLHE40 (rabbit polyclonal antibody; #ABN1737, Millipore-Sigma, Burlington, MA), and GAPDH (rabbit monoclonal antibody; #2118, Cell Signaling Technology, Danvers, MA, USA), were diluted in blocking solution at 4°C overnight.

Techniques: Expressing, Control, Plasmid Preparation

Journal: Oncology Research

Article Title: BHLHE40 Is a Transcriptional Regulatory Target of NFE2L3 in Triple-Negative Breast Cancer

doi: 10.32604/or.2025.070793

Figure Lengend Snippet: Validation of regulatory relationship between NFE2L3 and BHLHE40. ( A ) Visual representation of luciferase assay with BHLHE40 conjugated with Nanoluc vector, activated by NFE2L3, resulting in production of luciferase activity. Qualitative analysis of BHLHE40 transcriptional activation by NFE2L3 plasmid compared to BHLHE40 plasmid conjugated with the Nanoluc vector in ( B ) MDA-MB-31 and ( C ) MDA-MB-468 cells. Co-immunoprecipitation analysis of NFE2L3 protein and BHLHE40 protein interaction. ( D ) Co-IP of NFE2L3 pulldown with immunoblotting with BHLHE40. ( E ) Co-IP of BHLHE40 pulldown with immunoblotting with NFE2L3. Red boxes indicate the interacting bands. Student’s t -test was used to compare the two groups, *indicates p < 0.05

Article Snippet: The membranes were washed with PBST solution for 5 min three times, and then incubated with primary antibodies against NFE2L3 (rabbit polyclonal antibody; #A09888, Boster Bio, Pleasanton, CA, USA), BHLHE40 (rabbit polyclonal antibody; #ABN1737, Millipore-Sigma, Burlington, MA), and GAPDH (rabbit monoclonal antibody; #2118, Cell Signaling Technology, Danvers, MA, USA), were diluted in blocking solution at 4°C overnight.

Techniques: Biomarker Discovery, Luciferase, Plasmid Preparation, Activity Assay, Activation Assay, Immunoprecipitation, Co-Immunoprecipitation Assay, Western Blot

Journal: Oncology Research

Article Title: BHLHE40 Is a Transcriptional Regulatory Target of NFE2L3 in Triple-Negative Breast Cancer

doi: 10.32604/or.2025.070793

Figure Lengend Snippet: Phenotypic studies on MDA-MB-231 cells. Effect of knockdown or induction of NFE2L3 and BHLHE40 gene expression on MDA-MB-231 cell proliferation. MDA-MB-231 cell s’ proliferation was determined following treatment with vehicle (Control), siNFE2L3, siBHLHE40, as well as ( A ) NFE2L3 DNA plasmid, or ( B ) BHLHE40 DNA plasmid for 24, 48, and 72 h. Effect of knockdown or induction of NFE2L3 and BHLHE40 gene expression on MDA-MB-231 cell migration. MDA-MB-231 cell migration was determined following treatment with vehicle (Control), siNFE2L3, siBHLHE40, as well as ( C ) NFE2L3 DNA plasmid, or ( D ) BHLHE40 DNA plasmid for 24, 48, and 72 h. (Scale — 100 μm) ( E , F ) Graphical presentation of the estimation of wound closure area following treatment as described above. One-way ANOVA followed by post-hoc Tukey’s test was used. Compared to untreated: *indicates p < 0.05, **indicates p < 0.01, ***indicates p < 0.001, and ****indicates p < 0.0001. Compared to siNFE2L3: # indicates p < 0.05, ### indicates p < 0.001, and #### indicates p < 0.0001. Compared to siBHLHE40, a indicates p < 0.05, aaa indicates p < 0.001, and aaaa indicates p < 0.0001

Article Snippet: The membranes were washed with PBST solution for 5 min three times, and then incubated with primary antibodies against NFE2L3 (rabbit polyclonal antibody; #A09888, Boster Bio, Pleasanton, CA, USA), BHLHE40 (rabbit polyclonal antibody; #ABN1737, Millipore-Sigma, Burlington, MA), and GAPDH (rabbit monoclonal antibody; #2118, Cell Signaling Technology, Danvers, MA, USA), were diluted in blocking solution at 4°C overnight.

Techniques: Knockdown, Gene Expression, Control, Plasmid Preparation, Migration

Journal: Nature Communications

Article Title: Inability to switch from ARID1A-BAF to ARID1B-BAF impairs exit from pluripotency and commitment towards neural crest formation in ARID1B -related neurodevelopmental disorders

doi: 10.1038/s41467-021-26810-x

Figure Lengend Snippet: a , b CNCC differentiation was optimized using an ARID1B -wt control line. After 14 days, the cells exhibited the classic CNCC morphology and expressed the CNCC markers. P values are from two-sided Student’s t test. N = 3 biologically independent samples. Error bars display standard errors. c Time-course immunoblot conducted using Control Line-1 during CNCC differentiation shows that ARID1B is active in the first 7 days of the differentiation, with a peak of activity between day 5 and day 7. The ARID1B protein level strongly decreases after day 7. The experiment was repeated twice (the second replicate is shown in Fig. ). Marker bars display kDa. d , e Flow cytometry quantifying the expression of surface markers for pluripotency and CNCC differentiation in Control Line-1 and in the two patient lines. A large cell population is still pluripotent in both patients after 14 days ( d ). The patient lines also show reduced expression of CNCC surface markers after 14 days of differentiation relative to an ARID1B -wt control line at the same time point ( e ).

Article Snippet: CD99 Flow Cytometry: Miltenyi Biotech 130-121-086.

Techniques: Western Blot, Activity Assay, Marker, Flow Cytometry, Expressing

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Protective effects of Rosavin on bleomycin-induced pulmonary fibrosis via suppressing fibrotic and inflammatory signaling pathways in mice.

doi: 10.1016/j.biopha.2019.108870

Figure Lengend Snippet: Fig. 8. The effect of Rosavin on the expression of TGF-β1, α-SMA, Nrf2 and NF-κB p65 of BIM-induced PF in mice. Values are the mean ± SD (n = 3); nsP > 0.05, ##P < 0.01 vs. control group. *P < 0.05, **P < 0.01 vs. Bleomycin control group.

Article Snippet: Rabbit anti-TGF-β1 antibody (21898-1-AP) was purchased from Proteintech (Wuhan, China), and mouse anti-α-SMA antibody (BM0002), rabbit anti-NF-Kb p65 antibody, rabbit anti-Nrf2 antibody were all obtained from Wuhan Boster Biological Engineering Co., Ltd. (Wuhan, China).

Techniques: Expressing, Control

Journal: Chinese Medicine

Article Title: A clinical experience-based Chinese herbal formula improves ethanol-induced drunken behavior and hepatic steatohepatitis in mice models

doi: 10.1186/s13020-023-00753-5

Figure Lengend Snippet: The metabolic enzymes ADH1, ALDH2 and CYP2E1 are significantly up-regulated by alcohol, and further up-regulated by BGXJW administration. A – E The mRNA levels of ADH1, ALDH2, ALDH1, CYP2E1 in liver tissues were subjected to RT-qPCR analysis. E Levels of ADH1, ALDH2, CYP2E1 in liver lysates after indicated treatment were determined by western blot, F Liver tissue sections were subjected to immunohistochemistry analysis of CYP2E1. All data are expressed as the mean ± SEM (n = 3). nsP > 0.05, *P < 0.05 vs ctrl group; nsP > 0.05 vs EtOH group. Crtl contrl, EtOH ethanol model, BGXJW Bao-Gan-Xing-Jiu-Wan

Article Snippet: The antibodies used against CYP2E1, SCD1, FASN, and PPAR-α were purchased from Abcam (Massachusetts, USA); antibodies against ADH1 and ALDH2 were purchased from Boster (California, USA) and Cell Signaling Technology (Boston, USA) separately.

Techniques: Quantitative RT-PCR, Western Blot, Immunohistochemistry