Journal: Chromosoma

Article Title: FancJ (Brip1) loss-of-function allele results in spermatogonial cell depletion during embryogenesis and altered processing of crossover sites during meiotic prophase I in mice.

doi: 10.1007/s00412-015-0549-2

Figure Lengend Snippet: Fig. 7 Western blot analysis of whole testis lysates from Fancj+/+ and FancjGT/GT males reveals specific alterations in DNA MMR proteins and BLM. a–g Quantitation of protein levels for BRCA1, BLM, MLH1, MLH3, MSH4, MSH5, and MUS81 normalized to β-tubulin, showing significant increases in protein levels in FancjGT/GT (gray bars) compared to Fancj+/+ (black bars) testis extracts for all except BRCA1 and MUS81

Article Snippet: Primary-antibody incubation was performed for 12 h at 4 °C at 1:1000 dilution (antibodies used are: MUS81 (NBP1-32054 from Novus Biochemicals), MSH4 (ab95096 from Abcam), MSH5 (LS-C101911 from LsBio), MLH1 (550838 from BD pharmingen), β-Tubulin (T8328 from Sigma),β-Actin (A2228 from Sigma), BLM (a gift from Dr. Raimundo Freire), BRIP1/FANCJ (Novus, NBP1-31883), and BRCA1 (a gift from Dr. Satoshi Namekawa).

Techniques: Western Blot, Quantitation Assay

Journal: Communications Biology

Article Title: BHLHE40 contributes to allergic asthma progression in mice through NRTN downregulation in macrophages

doi: 10.1038/s42003-025-08288-1

Figure Lengend Snippet: A Schematic graph of animal sensitization and challenge. B Histological analysis of lung tissues stained with HE staining or PAS staining. C Histology score of HE staining and PAS staining. Statistical significance was determined with Mann-Whitney test. D mRNA expression of Bhlhe40 was assayed by RT-qPCR. Statistical analysis was performed by two-tailed Student’s t-test. E Western blot analysis of BHLBE40 in lung tissue. Statistical analysis was performed by two-tailed Student’s t-test. F Immunohistochemistry staining was performed to detected the level of BHLHE40 in lung tissue. Statistical analysis was performed by two-tailed Student’s t-test. G Double immunofluorescence staining for BHLHE40 (red) and F4/80 (green) in lung tissues. The number of double positive (F4/80 + BHLHE40 + ) cells was quantified. Statistical analysis was performed by two-tailed Student’s t-test. Error bars represent standard deviation. N = 8 biological replicates.

Article Snippet: For double immunofluorescence staining, slides were incubated with primary antibody against BHLHE40 (1:500, NB100-1800SS, Novus), and F4/80 (1:50, Sc-377009, Santa cruz) at 4 °C overnight.

Techniques: Staining, MANN-WHITNEY, Expressing, Quantitative RT-PCR, Two Tailed Test, Western Blot, Immunohistochemistry, Double Immunofluorescence Staining, Standard Deviation

Journal: Communications Biology

Article Title: BHLHE40 contributes to allergic asthma progression in mice through NRTN downregulation in macrophages

doi: 10.1038/s42003-025-08288-1

Figure Lengend Snippet: A Mice in asthma group received lentiviral delivery of LV- shBhlhe40 or LV-shNC. B RT-qPCR tested the mRNA expression of Bhlhe40 in lung tissues. Statistical analysis was performed by One-way ANOVA with Tukey’s post-test. C Representative western blot and quantitative analysis of BHLHE40 expression in lung tissues. Statistical analysis was performed by One-way ANOVA with Tukey’s post-test. D Representative HE images in lung tissue sections and quantitative analysis of histologic scoring. Statistical analysis was performed by Kruskal-Wallis test. E Representative PAS images in lung tissue sections and quantitative analysis of histologic scoring. Statistical analysis was performed by Kruskal-Wallis test. F Total protein concentrations in BALF in OVA-induced asthma. Statistical analysis was performed by One-way ANOVA with Tukey’s post-test. Error bars represent standard deviation. N = 8 biological replicates.

Article Snippet: For double immunofluorescence staining, slides were incubated with primary antibody against BHLHE40 (1:500, NB100-1800SS, Novus), and F4/80 (1:50, Sc-377009, Santa cruz) at 4 °C overnight.

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Standard Deviation

Journal: Communications Biology

Article Title: BHLHE40 contributes to allergic asthma progression in mice through NRTN downregulation in macrophages

doi: 10.1038/s42003-025-08288-1

Figure Lengend Snippet: A ELISA forYM1 in BALF. B – E RT-qPCR assay showed the mRNA expression of Ym1 , Fizz1 , Cd206 , Arg1 in lung tissues. F The protein level of CD206 and ARG1 in lung tissues. G Quantitative analysis of CD206 protein level. H Quantitative analysis of ARG1 protein level. I Western blot assessed the level of BHLHE40, CD206, and ARG1 in monocyte-derived alveolar macrophages (Mo-AMs). J Quantitative analysis of BHLHE40 protein level in Mo-AMs. K Quantitative analysis of CD206 protein level in Mo-AMs. L Quantitative analysis of ARG1 protein level in Mo-AMs. N = 8 biological replicates. Statistical analysis was performed by One-way ANOVA with Tukey’s post-test. Error bars represent standard deviation.

Article Snippet: For double immunofluorescence staining, slides were incubated with primary antibody against BHLHE40 (1:500, NB100-1800SS, Novus), and F4/80 (1:50, Sc-377009, Santa cruz) at 4 °C overnight.

Techniques: Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Expressing, Western Blot, Derivative Assay, Standard Deviation

Journal: Communications Biology

Article Title: BHLHE40 contributes to allergic asthma progression in mice through NRTN downregulation in macrophages

doi: 10.1038/s42003-025-08288-1

Figure Lengend Snippet: A Bone marrow derived macrophages (BMDMs) were isolated from BALB/c mice, and then stimulated with IL-4 to induce M1 polarization. Lentivirus infection was conducted before IL-4 treatment. B The expression level of Bhlhe40 was determined by RT-qPCR. C Western blot analysis of BHLBE40 in BMDMs. D – G The mRNA expression level of Ym1 , Fizz1 , Cd206 , and Arg1 in BMDMs. H The protein level of CD206 and ARG1 in BMDMs. I Immunofluorescence analysis of ARG1 expression in BMDMs, and the quantification of ARG1 positive cell count. N = 4 biological replicates. Statistical analysis was performed by One-way ANOVA with Tukey’s post-test. Error bars represent standard deviation.

Article Snippet: For double immunofluorescence staining, slides were incubated with primary antibody against BHLHE40 (1:500, NB100-1800SS, Novus), and F4/80 (1:50, Sc-377009, Santa cruz) at 4 °C overnight.

Techniques: Derivative Assay, Isolation, Infection, Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Cell Counting, Standard Deviation

Journal: Communications Biology

Article Title: BHLHE40 contributes to allergic asthma progression in mice through NRTN downregulation in macrophages

doi: 10.1038/s42003-025-08288-1

Figure Lengend Snippet: A , B Nrtn expression level was determined by RT-qPCR and western blot. Statistical analysis was performed by One-way ANOVA with Tukey’s post-test. C Dual-luciferase reporter assay. N = 3 biological replicates. Statistical analysis was performed by One-way ANOVA with Tukey’s post-test. D Validation by ChIP-qPCR analysis of BHLHE40 binding to Nrtn in BMDMs. Statistical analysis was performed by two-tailed Student’s t-test. E RT-qPCR showed the mRNA expression of Fizz1 , Ym1 , Arg1 , and Cd206 . Statistical analysis was performed by One-way ANOVA with Tukey’s post-test. F The protein level of CD206 and ARG1 was assayed by western blot. Statistical analysis was performed by One-way ANOVA with Tukey’s post-test. G Immunofluorescence analysis of ARG1 expression in BMDMs, and the quantification of ARG1 positive cell count. Statistical analysis was performed by One-way ANOVA with Tukey’s post-test. Error bars represent standard deviation. N = 3-4 biological replicates.

Article Snippet: For double immunofluorescence staining, slides were incubated with primary antibody against BHLHE40 (1:500, NB100-1800SS, Novus), and F4/80 (1:50, Sc-377009, Santa cruz) at 4 °C overnight.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Luciferase, Reporter Assay, Biomarker Discovery, ChIP-qPCR, Binding Assay, Two Tailed Test, Immunofluorescence, Cell Counting, Standard Deviation

Journal: Communications Biology

Article Title: BHLHE40 contributes to allergic asthma progression in mice through NRTN downregulation in macrophages

doi: 10.1038/s42003-025-08288-1

Figure Lengend Snippet: A Mice were stimulated with thioglycolate and IL-4 by intraperitoneal injection, and then physiological peritoneal macrophages were obtained. B Validation by ChIP-qPCR analysis of BHLHE40 binding to Nrtn in peritoneal macrophages. Statistical analysis was performed by two-tailed Student’s t-test. C RT-qPCR examined the mRNA level of Bhlhe40 in peritoneal macrophages. Statistical analysis was performed by One-way ANOVA with Tukey’s post-test. D Representative immunoblot and quantitative analysis of BHLHE40 expression. Statistical analysis was performed by One-way ANOVA with Tukey’s post-test. E Immunofluorescence analysis of ARG1 expression in peritoneal macrophages, and the quantification of ARG1 positive cell count. Statistical analysis was performed by One-way ANOVA with Tukey’s post-test. F Peritoneal macrophages were infected with LV- shBhlhe40 and LV- shNrtn , and then were subjected to immunofluorescence staining for ARG1. The number of ARG1 positive cells was quantified. Statistical analysis was performed by One-way ANOVA with Tukey’s post-test. Error bars represent standard deviation. N = 4 biological replicates.

Article Snippet: For double immunofluorescence staining, slides were incubated with primary antibody against BHLHE40 (1:500, NB100-1800SS, Novus), and F4/80 (1:50, Sc-377009, Santa cruz) at 4 °C overnight.

Techniques: Injection, Biomarker Discovery, ChIP-qPCR, Binding Assay, Two Tailed Test, Quantitative RT-PCR, Western Blot, Expressing, Immunofluorescence, Cell Counting, Infection, Staining, Standard Deviation