Journal: International Journal of Molecular Sciences

Article Title: 5-Methylcytosine and 5-Hydroxymethylcytosine in Scrapie-Infected Sheep and Mouse Brain Tissues

doi: 10.3390/ijms24021621

Figure Lengend Snippet: Correlations in the thalamus region of 5mC and 5hmC with the expression levels of some genes involved in epigenetic regulation. 5hmC positively correlated with ( a ) Dnmt1 , ( b ) Dnmt3a , ( c ) Tet1 , ( d ) Tet2 , and ( e ) Hdac1 in Tg338 mice, and 5mC ( f ) and 5hmC ( g ) negatively correlated with HDAC2 in sheep.

Article Snippet: Hdac1 , Mm02745760_g1.

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: 5-Methylcytosine and 5-Hydroxymethylcytosine in Scrapie-Infected Sheep and Mouse Brain Tissues

doi: 10.3390/ijms24021621

Figure Lengend Snippet: References of the probes used for the RT-qPCR TaqMan amplification assay in Tg338 mice. ID = identification of the commercial company (Thermo Fisher Scientific, Waltham, MA, USA).

Article Snippet: Hdac1 , Mm02745760_g1.

Techniques: Amplification

Journal: International Journal of Molecular Sciences

Article Title: 5-Methylcytosine and 5-Hydroxymethylcytosine in Scrapie-Infected Sheep and Mouse Brain Tissues

doi: 10.3390/ijms24021621

Figure Lengend Snippet: Sequences of primers used in the RT-qPCR assay in sheep. Fw = Forward primer and Rv = Reverse primer.

Article Snippet: Hdac1 , Mm02745760_g1.

Techniques:

Journal: Inflammation Research

Article Title: SDS3 regulates microglial inflammation by modulating the expression of the upstream kinase ASK1 in the p38 MAPK signaling pathway

doi: 10.1007/s00011-024-01913-5

Figure Lengend Snippet: Knockdown of SDS3 enhances lipopolysaccharide (LPS)-induced inflammatory responses in microglia. ( A ) Immunoprecipitation (IP) followed by Western blotting confirmed the interaction between SDS3, Sin3A, and HDAC1 in BV2 cells. ( B ) Protein levels of SDS3, Sin3A, and HDAC1 in BV2 cells after treatment with 1 μg/mL LPS for different lengths of time. β-Actin was used as the loading control. ( C ) Protein levels of SDS3, Sin3A, and HDAC1 in BV2 cells after treatment with different concentrations of LPS for 6 h. β-Actin was used as the loading control. ( D ) Changes in SDS3 mRNA expression levels in BV2 cells after treatment with 1 μg/mL LPS for different lengths of time. ACTB was used as the reference gene. The control group represents 0 h; n = 3; error bars represent mean ± standard error of the mean (SEM). Data were analyzed by one-way analysis of variance; * p < 0.05. ( E ) Protein levels of SDS3 in BV2 cells after treatment with 4 μM MG132 or DMSO for 1 h, followed by treatment with 1 μg/mL LPS for 6 h. β-Actin was used as the loading control. ( F ) Protein levels of iNOS, COX-2, and IL-1β in BV2 cells transfected with SDS3 or control siRNA, followed by treatment with 1 μg/mL LPS for 6 h. β-Actin was used as the loading control. ( G ) mRNA levels of SDS3, iNOS, COX-2, and IL-1β in BV2 cells transfected with SDS3 or Control siRNA, followed by treatment with 1 μg/mL LPS for 6 h. ACTB was used as the reference gene. The Control siRNA group that was not treated with LPS served as the control; n = 3. Error bars represent mean ± SEM. Data were analyzed by Student’s t-test; * p < 0.01; ** p < 0.01; *** p < 0.001. ( H ) Concentrations of nitrite as a measure of nitric oxide production in the culture medium of BV2 cells transfected with SDS3 or Control siRNA, followed by treatment with 1 μg/mL LPS for 24 h, were determined using the Griess Reagent method; n = 3. Error bars represent mean ± SEM. Statistical analysis was performed using one-way analysis of variance followed by a Tukey’s multiple comparisons test; * p < 0.01. For protein band quantification information, please refer to Supplemental Fig.

Article Snippet: Control siRNA (sc-37,007), SDS3 siRNA (sc-153,291), and HDAC1 siRNA (sc-29,344) were obtained from Santa Cruz and were transfected into BV2 cells using Lipofectamine RNAiMAX (Invitrogen, 13,778,030).

Techniques: Knockdown, Immunoprecipitation, Western Blot, Control, Expressing, Transfection

Journal: Inflammation Research

Article Title: SDS3 regulates microglial inflammation by modulating the expression of the upstream kinase ASK1 in the p38 MAPK signaling pathway

doi: 10.1007/s00011-024-01913-5

Figure Lengend Snippet: Regulation of ASK1 expression by SDS3 and HDAC1. ( A ) Binding of SDS3 to the promoter region of ASK1 . ChIP experiments were performed using anti-SDS3 or negative-control IgG on chromatin fractions from formaldehyde cross-linked BV2 cells. Quantitative PCR (qPCR) analysis was conducted to assess the binding of SDS3 to the promoter region of ASK1 . Results are presented as a percentage of input. IgG was used as the control group; n = 3. Error bars represent mean ± SEM. Data were analyzed by Student’s t-test; ** p < 0.01. ( B ) Upregulation of ASK1 expression after SDS3 knockdown. BV2 cells were transfected with SDS3 or Control siRNA, and changes in SDS3 and ASK1 mRNA and protein expression were measured. β-Actin was used as the internal reference. The Control siRNA group served as the control group; n = 3. Error bars represent mean ± SEM. Data were analyzed by Student’s t-test; * p < 0.05; ** p < 0.01; *** p < 0.001. ( C ) Binding of HDAC1 to the ASK1 promoter. ChIP experiments were performed using anti-HDAC1 or negative-control IgG on chromatin fractions from formaldehyde cross-linked BV2 cells. qPCR analysis was conducted to assess the binding of HDAC1 to the ASK1 promoter region. Results are presented as a percentage of input. IgG was used as the control group; n = 3. Error bars represent mean ± SEM. Data were analyzed by Student’s t-test; * p < 0.05; ** p < 0.01. (D) Upregulation of ASK1 expression after HDAC1 knockdown. BV2 cells were transfected with HDAC1 or Control siRNA, and changes in HDAC1 and ASK1 mRNA and protein expression were measured. β-Actin was used as the internal reference. The Control siRNA group served as the control group; n = 3. Error bars represent mean ± SEM. Data were analyzed by Student’s t-test; * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: Control siRNA (sc-37,007), SDS3 siRNA (sc-153,291), and HDAC1 siRNA (sc-29,344) were obtained from Santa Cruz and were transfected into BV2 cells using Lipofectamine RNAiMAX (Invitrogen, 13,778,030).

Techniques: Expressing, Binding Assay, Negative Control, Real-time Polymerase Chain Reaction, Control, Knockdown, Transfection