stat5b  (Proteintech)

Journal: Cells

Article Title: Oligodendrocyte-Specific STAT5B Overexpression Ameliorates Myelin Impairment in Experimental Models of Parkinson’s Disease

doi: 10.3390/cells14151145

Figure Lengend Snippet: Discovery and validation of STAT5B as a key regulatory factor. ( A ) DoRothEA analysis identifies 32 TFs with significantly altered activity in oligodendrocytes in the PD group (12 activated, 20 downregulated). ( B ) Differential ranking of TFs reveals significant downregulation of STAT5B mRNA in snRNA-seq results (****, p < 0.0001). ( C ) Violin plot confirms significant decrease in STAT5B expression in oligodendrocytes from the PD group compared to the control group. ( D , E ) LFB staining shows decreased myelin density correlating with STAT5B mRNA (r = 0.83, p < 0.00074) and protein (r = 0.76, p < 0.0039) expression in the SN of mice.

Article Snippet: Primary antibody incubations (4 °C overnight) included the following: Olig2 mouse monoclonal (1:300, sc-515947, Santa Cruz Biotechnology, Shanghai, China), STAT5B rabbit (1:200, CSB-PA022815LA01HU, CUSABIO, Wuhan, China) and mouse (1:200, 66427-1-Ig, Proteintech, Wuhan, China) monoclonals, and MBP rabbit monoclonal (1:200, 10458-1-AP, Proteintech, Wuhan, China).

Techniques: Biomarker Discovery, Activity Assay, Expressing, Control, Staining

Journal: Cells

Article Title: Oligodendrocyte-Specific STAT5B Overexpression Ameliorates Myelin Impairment in Experimental Models of Parkinson’s Disease

doi: 10.3390/cells14151145

Figure Lengend Snippet: The expression of STAT5B was significantly reduced in the PD model. ( A ) Representative image of immunofluorescence staining in the SN of the MPTP-induced mouse model (scale bars = 100 μm). ( B ) Quantification of co-localization using Pearson’s correlation coefficient (PCC). The y-axis represents the PCC, which quantifies the degree of spatial overlap between the STAT5B and Olig2 signals. PCC values range from −1 to +1, where +1 indicates perfect co-localization, 0 indicates no correlation (random distribution), and -1 indicates perfect segregation. Each data point represents the PCC calculated from individual image fields. The data are presented as the means ± SEMs of 3 independent experiments. Data are presented from male (n = 3) mice, aged 8 weeks. ( C ) qRT-PCR analysis of STAT5B mRNA expression in MO3.13 cells treated with 200 and 500 μM MPP + for 24 h. The data are presented as the means ± SEMs of 3 independent experiments. ( D , E ) Western blot analysis of STAT5B protein expression in MO3.13 cells treated with 500 μM MPP + for 24 h. The data are presented as the means ± SEMs of 6 independent experiments (*, p < 0.05; **, p < 0.01).

Article Snippet: Primary antibody incubations (4 °C overnight) included the following: Olig2 mouse monoclonal (1:300, sc-515947, Santa Cruz Biotechnology, Shanghai, China), STAT5B rabbit (1:200, CSB-PA022815LA01HU, CUSABIO, Wuhan, China) and mouse (1:200, 66427-1-Ig, Proteintech, Wuhan, China) monoclonals, and MBP rabbit monoclonal (1:200, 10458-1-AP, Proteintech, Wuhan, China).

Techniques: Expressing, Immunofluorescence, Staining, Quantitative RT-PCR, Western Blot

Journal: Cells

Article Title: Oligodendrocyte-Specific STAT5B Overexpression Ameliorates Myelin Impairment in Experimental Models of Parkinson’s Disease

doi: 10.3390/cells14151145

Figure Lengend Snippet: LFB staining of myelin in co-cultures of differentiated MO3.13 cells and SH-SY5Y neuronally differentiated cells. ( A , B ) LFB staining of STAT5B overexpression in MPP + -treated MO3.13 cells. ( C , D ) LFB staining of STAT5B knockdown in MO3.13 cells. The results are expressed as the mean ± SD (n = 3; 10× scale bars = 200 μm; **, p < 0.01; ***, p < 0.001).

Article Snippet: Primary antibody incubations (4 °C overnight) included the following: Olig2 mouse monoclonal (1:300, sc-515947, Santa Cruz Biotechnology, Shanghai, China), STAT5B rabbit (1:200, CSB-PA022815LA01HU, CUSABIO, Wuhan, China) and mouse (1:200, 66427-1-Ig, Proteintech, Wuhan, China) monoclonals, and MBP rabbit monoclonal (1:200, 10458-1-AP, Proteintech, Wuhan, China).

Techniques: Staining, Over Expression, Knockdown

Journal: Cells

Article Title: Oligodendrocyte-Specific STAT5B Overexpression Ameliorates Myelin Impairment in Experimental Models of Parkinson’s Disease

doi: 10.3390/cells14151145

Figure Lengend Snippet: Overexpression of oligodendrocyte STAT5B improved myelin damage in MPTP-induced mice. ( A , B ) LFB staining showed that overexpression of oligodendrocyte STAT5B increased myelin density in MPTP-induced mice. The results are expressed as the mean ± SD (10× scale bars = 200 μm). Data are presented from male (n = 6) mice, aged 8 weeks. ( C , D ) TEM analysis revealed that oligodendrocyte STAT5B overexpression increased axonal myelin thickness and significantly reduced g-ratio in MPTP-treated mice. The results are expressed as the mean ± SD (5 randomly selected myelinated axons per SN field; 20.0 k× scale bars = 1 μm). ( E ) Scatter plot illustrating the individual g -ratio values and the distribution of axonal sizes. Data are presented from male (n = 3) mice, aged 8 weeks (*, p < 0.05; **, p < 0.01; ***, p < 0.01).

Article Snippet: Primary antibody incubations (4 °C overnight) included the following: Olig2 mouse monoclonal (1:300, sc-515947, Santa Cruz Biotechnology, Shanghai, China), STAT5B rabbit (1:200, CSB-PA022815LA01HU, CUSABIO, Wuhan, China) and mouse (1:200, 66427-1-Ig, Proteintech, Wuhan, China) monoclonals, and MBP rabbit monoclonal (1:200, 10458-1-AP, Proteintech, Wuhan, China).

Techniques: Over Expression, Staining

Journal: Cells

Article Title: Oligodendrocyte-Specific STAT5B Overexpression Ameliorates Myelin Impairment in Experimental Models of Parkinson’s Disease

doi: 10.3390/cells14151145

Figure Lengend Snippet: Overexpression of oligodendrocyte STAT5B ameliorated dopamine neuronal damage in MPTP-treated mice. ( A ) qRT-PCR analysis of TH mRNA expression in the mouse SN. ( B ) Western blot analysis and ( C ) quantification of TH protein levels in the mouse SN. ( D , E ) Immunohistochemical staining of TH protein in the mouse SN. ( F , G ) Immunohistochemical staining of NfL protein in the mouse SN. The results are expressed as the mean ± SD (10× scale bars = 200 μm; 40× scale bars = 50 μm; *, p < 0.05; **, p < 0.01; ***, p < 0.001). Data are presented from male (n = 6) mice, aged 8 weeks.

Article Snippet: Primary antibody incubations (4 °C overnight) included the following: Olig2 mouse monoclonal (1:300, sc-515947, Santa Cruz Biotechnology, Shanghai, China), STAT5B rabbit (1:200, CSB-PA022815LA01HU, CUSABIO, Wuhan, China) and mouse (1:200, 66427-1-Ig, Proteintech, Wuhan, China) monoclonals, and MBP rabbit monoclonal (1:200, 10458-1-AP, Proteintech, Wuhan, China).

Techniques: Over Expression, Quantitative RT-PCR, Expressing, Western Blot, Immunohistochemical staining, Staining

Journal: Cells

Article Title: Oligodendrocyte-Specific STAT5B Overexpression Ameliorates Myelin Impairment in Experimental Models of Parkinson’s Disease

doi: 10.3390/cells14151145

Figure Lengend Snippet: Overexpression of oligodendrocyte STAT5B improved motor function in MPTP-treated mice. ( A , B ) Pole test results showing total time and turn time. ( C ) Rotarod test results showing fall latency. ( D – F ) Gait analysis results showing movement speed, stride length, and support time. NS: no support, SLS: single-leg support, CLS: contralateral limb support, HLS: homologous limb support, ILS: ipsilateral limb support, TLS: three-limb support, FLS: four-limb support. The results are expressed as the mean ± SD (*, p < 0.05; **, p < 0.01; ***, p < 0.001). Data are presented from male (n = 14) mice, aged 8 weeks.

Article Snippet: Primary antibody incubations (4 °C overnight) included the following: Olig2 mouse monoclonal (1:300, sc-515947, Santa Cruz Biotechnology, Shanghai, China), STAT5B rabbit (1:200, CSB-PA022815LA01HU, CUSABIO, Wuhan, China) and mouse (1:200, 66427-1-Ig, Proteintech, Wuhan, China) monoclonals, and MBP rabbit monoclonal (1:200, 10458-1-AP, Proteintech, Wuhan, China).

Techniques: Over Expression

Journal: Cells

Article Title: Oligodendrocyte-Specific STAT5B Overexpression Ameliorates Myelin Impairment in Experimental Models of Parkinson’s Disease

doi: 10.3390/cells14151145

Figure Lengend Snippet: STAT5B overexpression promoted MBP expression and reduced myelin injury. ( A , B ) Luciferase reporter assay showing the binding efficiency of STAT5B to the MBP promoter region (n = 6). ( C – H ) In MO3.13 cells overexpressing STAT5B : ( C ) qRT-PCR and ( D , E ) Western blot analysis of MBP expression (n = 4). ( F – H ) Immunofluorescently stained STAT5B and MBP expression (n = 3, scale bars = 50 μm). The results are expressed as the mean ± SD (**, p < 0.01; ***, p < 0.001).

Article Snippet: Primary antibody incubations (4 °C overnight) included the following: Olig2 mouse monoclonal (1:300, sc-515947, Santa Cruz Biotechnology, Shanghai, China), STAT5B rabbit (1:200, CSB-PA022815LA01HU, CUSABIO, Wuhan, China) and mouse (1:200, 66427-1-Ig, Proteintech, Wuhan, China) monoclonals, and MBP rabbit monoclonal (1:200, 10458-1-AP, Proteintech, Wuhan, China).

Techniques: Over Expression, Expressing, Luciferase, Reporter Assay, Binding Assay, Quantitative RT-PCR, Western Blot, Staining

Journal: Cells

Article Title: Oligodendrocyte-Specific STAT5B Overexpression Ameliorates Myelin Impairment in Experimental Models of Parkinson’s Disease

doi: 10.3390/cells14151145

Figure Lengend Snippet: Knockdown of STAT5B decreases MBP expression and aggravates myelin damage. ( A – F ) In STAT5B -knockdown MO3.13 cells: ( A ) qRT-PCR analysis of MBP mRNA expression (n = 4), ( B , C ) Western blot analysis of MBP protein expression (n = 4), and ( D – F ) immunofluorescently stained STAT5B and MBP expression (n = 3, scale bars = 50 μm). ( G ) qRT-PCR analysis of MBP mRNA expression in the mouse SN. ( H , I ) Western blot analysis and quantification of MBP protein levels in the mouse SN. The results are expressed as the mean ± SD (*, p < 0.05; **, p < 0.01; ***, p < 0.001). Data are presented from male (n = 6) mice, aged 8 weeks.

Article Snippet: Primary antibody incubations (4 °C overnight) included the following: Olig2 mouse monoclonal (1:300, sc-515947, Santa Cruz Biotechnology, Shanghai, China), STAT5B rabbit (1:200, CSB-PA022815LA01HU, CUSABIO, Wuhan, China) and mouse (1:200, 66427-1-Ig, Proteintech, Wuhan, China) monoclonals, and MBP rabbit monoclonal (1:200, 10458-1-AP, Proteintech, Wuhan, China).

Techniques: Knockdown, Expressing, Quantitative RT-PCR, Western Blot, Staining

Journal: Cells

Article Title: Oligodendrocyte-Specific STAT5B Overexpression Ameliorates Myelin Impairment in Experimental Models of Parkinson’s Disease

doi: 10.3390/cells14151145

Figure Lengend Snippet: STAT5B mRNA stability and promoter methylation study. ( A ) Actinomycin D transcription inhibition assay to detect STAT5B mRNA stability. Line chart showing STAT5B mRNA expression at different time points after treatment with actinomycin D. The results are expressed as the mean ± SD (n = 6). ( B ) The MethPrimer website predicts that there are extensive CpG islands in the promoter region of STAT5B (light blue areas are CpG island regions). ( C , D ) Top 10 CpG sites with significantly increased methylation in the PD group compared to the control group, as determined by MethylTarget sequencing: ( C ) MethylTarget sequencing scatter plot—Y-axis: “CG methyl level”, “CG density”; X-axis: methylation site positions. ( D ) MethylTarget sequencing heatmap—Y-axis: methylation sites; X-axis: groups.

Article Snippet: Primary antibody incubations (4 °C overnight) included the following: Olig2 mouse monoclonal (1:300, sc-515947, Santa Cruz Biotechnology, Shanghai, China), STAT5B rabbit (1:200, CSB-PA022815LA01HU, CUSABIO, Wuhan, China) and mouse (1:200, 66427-1-Ig, Proteintech, Wuhan, China) monoclonals, and MBP rabbit monoclonal (1:200, 10458-1-AP, Proteintech, Wuhan, China).

Techniques: Methylation, Inhibition, Expressing, Control, Sequencing

Journal: Cells

Article Title: Oligodendrocyte-Specific STAT5B Overexpression Ameliorates Myelin Impairment in Experimental Models of Parkinson’s Disease

doi: 10.3390/cells14151145

Figure Lengend Snippet: Effects of DNMT3A on methylation levels in the STAT5B promoter region. ( A ) mRNA expression of DNA methylation-related genes assessed by qRT-PCR (n = 6, **, p < 0.01; ***, p < 0.001). ( B , C ) Methylation-specific PCR (MSP) analysis of the STAT5B promoter region at site 2202 in DNMT3A -knockdown and MPP + -induced MO3.13 cells. ( D , E ) MSP analysis of the STAT5B promoter region at site 2202 in DNMT3A -overexpression MO3.13 cells.

Article Snippet: Primary antibody incubations (4 °C overnight) included the following: Olig2 mouse monoclonal (1:300, sc-515947, Santa Cruz Biotechnology, Shanghai, China), STAT5B rabbit (1:200, CSB-PA022815LA01HU, CUSABIO, Wuhan, China) and mouse (1:200, 66427-1-Ig, Proteintech, Wuhan, China) monoclonals, and MBP rabbit monoclonal (1:200, 10458-1-AP, Proteintech, Wuhan, China).

Techniques: Methylation, Expressing, DNA Methylation Assay, Quantitative RT-PCR, Knockdown, Over Expression

Journal: Cells

Article Title: Oligodendrocyte-Specific STAT5B Overexpression Ameliorates Myelin Impairment in Experimental Models of Parkinson’s Disease

doi: 10.3390/cells14151145

Figure Lengend Snippet: DNMT3A knockdown and overexpression affect STAT5B expression. ( A – D ) In DNMT3A -knockdown MO3.13 cells: ( A – D ) qRT-PCR and ( C , D ) Western blot analysis of STAT5B expression (n = 6). ( E – H ) In MO3.13 cells with DNMT3A overexpression or STAT5B co-overexpression with DNMT3A : ( E , F ) qRT-PCR analysis of DNMT3A and STAT5B mRNA and ( G , H ) Western blot analysis of STAT5B protein expression (n = 6). The results are expressed as the mean ± SD (*, p < 0.05; **, p < 0.01; ***, p < 0.001).

Article Snippet: Primary antibody incubations (4 °C overnight) included the following: Olig2 mouse monoclonal (1:300, sc-515947, Santa Cruz Biotechnology, Shanghai, China), STAT5B rabbit (1:200, CSB-PA022815LA01HU, CUSABIO, Wuhan, China) and mouse (1:200, 66427-1-Ig, Proteintech, Wuhan, China) monoclonals, and MBP rabbit monoclonal (1:200, 10458-1-AP, Proteintech, Wuhan, China).

Techniques: Knockdown, Over Expression, Expressing, Quantitative RT-PCR, Western Blot

Journal: Cells

Article Title: Oligodendrocyte-Specific STAT5B Overexpression Ameliorates Myelin Impairment in Experimental Models of Parkinson’s Disease

doi: 10.3390/cells14151145

Figure Lengend Snippet: DNMT3A-mediated downregulation of MBP via STAT5B in oligodendrocytes. ( A – E ) In DNMT3A -knockdown MO3.13 cells: ( A – C ) qRT-PCR and Western blot analysis of MBP expression (n = 6). ( D , E ) LFB staining indicates improved myelin integrity in SH-SY5Y neuronally differentiated cells co-cultured with differentiated DNMT3A -knockdown MO3.13 cells (n = 3, scale bars = 200 μm). The results are expressed as the mean ± SD (*, p < 0.05; **, p < 0.01; ***, p < 0.001). ( F – J ) In MO3.13 cells with DNMT3A and STAT5B co-overexpression: ( F – H ) qRT-PCR and Western blot analysis of MBP expression (n = 6). ( I , J ) LFB staining shows improved myelin integrity in SH-SY5Y neuronally differentiated cells co-cultured with MO3.13 cells co-overexpressing STAT5B and DNMT3A (n = 3). The results are expressed as the mean ± SD (*, p < 0.05; **, p < 0.01; ***, p < 0.001).

Article Snippet: Primary antibody incubations (4 °C overnight) included the following: Olig2 mouse monoclonal (1:300, sc-515947, Santa Cruz Biotechnology, Shanghai, China), STAT5B rabbit (1:200, CSB-PA022815LA01HU, CUSABIO, Wuhan, China) and mouse (1:200, 66427-1-Ig, Proteintech, Wuhan, China) monoclonals, and MBP rabbit monoclonal (1:200, 10458-1-AP, Proteintech, Wuhan, China).

Techniques: Knockdown, Quantitative RT-PCR, Western Blot, Expressing, Staining, Cell Culture, Over Expression