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Addgene inc mbd3 overexpression

Fig. 3 Knockdown of <t>MBD3</t> promoted PKCi-derived mES self-renewal. A qPCR showed that mRNA levels of MBD3 decreased after PKCi- derived mES at passage 5 were transfected with shMBD3, with PKCi and shNC groups used as controls. B MBD3 and β-actin protein levels were evaluated by Western blot in PKCi-derived mES with MBD3 knockdown (upper panel). Quantitative density analysis showed that shMBD3 decreased protein levels of MBD3 (lower panel). C qPCR showed that compared with the control groups, shMBD3 increased the mRNA levels of pluripotency genes NANOG and OCT4 (left panel) but did not affect the mRNA levels of differentiation genes (right panel). D Western blot detection of NANOG, OCT4, and β-actin protein expression in PKCi-derived mES with MBD3 knockdown (left panel). Quantitative density analysis showed that shMBD3 increased protein levels of NANOG and OCT4 (right panel). E Knockdown of MBD3 did not affect the morphology or AP staining of PKCi-derived mES. Scale bar, 200 μm (upper panel). Knockdown of MBD3 did not affect the total number of AP- positive colonies (left panel) but increased the percentage of mixed colonies and decreased the percentage of differentiated colonies (right panel). Data were shown as mean ± SD (n = 3). The letters a and b indicated signiﬁcant differences among groups (P < 0.05).

Mbd3 Overexpression, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 93 stars, based on 2 article reviews

mbd3 overexpression - by Bioz Stars, 2026-04

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1) Product Images from "Nucleosome remodeling and deacetylation complex and MBD3 influence mouse embryonic stem cell naïve pluripotency under inhibition of protein kinase C."

Article Title: Nucleosome remodeling and deacetylation complex and MBD3 influence mouse embryonic stem cell naïve pluripotency under inhibition of protein kinase C.

Journal: Cell death discovery

doi: 10.1038/s41420-022-01131-0

Fig. 3 Knockdown of MBD3 promoted PKCi-derived mES self-renewal. A qPCR showed that mRNA levels of MBD3 decreased after PKCi- derived mES at passage 5 were transfected with shMBD3, with PKCi and shNC groups used as controls. B MBD3 and β-actin protein levels were evaluated by Western blot in PKCi-derived mES with MBD3 knockdown (upper panel). Quantitative density analysis showed that shMBD3 decreased protein levels of MBD3 (lower panel). C qPCR showed that compared with the control groups, shMBD3 increased the mRNA levels of pluripotency genes NANOG and OCT4 (left panel) but did not affect the mRNA levels of differentiation genes (right panel). D Western blot detection of NANOG, OCT4, and β-actin protein expression in PKCi-derived mES with MBD3 knockdown (left panel). Quantitative density analysis showed that shMBD3 increased protein levels of NANOG and OCT4 (right panel). E Knockdown of MBD3 did not affect the morphology or AP staining of PKCi-derived mES. Scale bar, 200 μm (upper panel). Knockdown of MBD3 did not affect the total number of AP- positive colonies (left panel) but increased the percentage of mixed colonies and decreased the percentage of differentiated colonies (right panel). Data were shown as mean ± SD (n = 3). The letters a and b indicated signiﬁcant differences among groups (P < 0.05).

Figure Legend Snippet: Fig. 3 Knockdown of MBD3 promoted PKCi-derived mES self-renewal. A qPCR showed that mRNA levels of MBD3 decreased after PKCi- derived mES at passage 5 were transfected with shMBD3, with PKCi and shNC groups used as controls. B MBD3 and β-actin protein levels were evaluated by Western blot in PKCi-derived mES with MBD3 knockdown (upper panel). Quantitative density analysis showed that shMBD3 decreased protein levels of MBD3 (lower panel). C qPCR showed that compared with the control groups, shMBD3 increased the mRNA levels of pluripotency genes NANOG and OCT4 (left panel) but did not affect the mRNA levels of differentiation genes (right panel). D Western blot detection of NANOG, OCT4, and β-actin protein expression in PKCi-derived mES with MBD3 knockdown (left panel). Quantitative density analysis showed that shMBD3 increased protein levels of NANOG and OCT4 (right panel). E Knockdown of MBD3 did not affect the morphology or AP staining of PKCi-derived mES. Scale bar, 200 μm (upper panel). Knockdown of MBD3 did not affect the total number of AP- positive colonies (left panel) but increased the percentage of mixed colonies and decreased the percentage of differentiated colonies (right panel). Data were shown as mean ± SD (n = 3). The letters a and b indicated signiﬁcant differences among groups (P < 0.05).

Techniques Used: Knockdown, Derivative Assay, Transfection, Western Blot, Control, Expressing, Staining

Fig. 4 Overexpression of MBD3-induced PKCi-derived mES differentiation. A qPCR showed that mRNA levels of MBD3 increased after PKCi- derived mES at passage 5 were transfected with FUW-MBD3, with the PKCi and FUW-M2rtTA plasmid groups used as controls. B MBD3 and β-actin protein levels were evaluated by Western blot after PKCi-derived mES were transfected with FUW-MBD3 (upper panel). Quantitative density analysis showed that protein levels of MBD3 increased after transfection with FUW-MBD3 (lower panel). C qPCR showed decreased mRNA levels of pluripotency genes NANOG and OCT4 and naïve-state marker REX1, but increased mRNA levels of primed-state marker FGF5 after MBD3 was overexpressed (left panel). MBD3 overexpression also increased mRNA levels of the endoderm marker CK8, mesoderm markers cTnT, BMP4, and DESMIN, and ectoderm markers PAX6 and SOX17 (right panel). D Western blot detection of NANOG, OCT4, REX1, FGF5, cTnT, and β-actin protein expression after MBD3 overexpression (left panel). Quantitative density analysis showed that MBD3 overexpression decreased protein levels of NANOG, OCT4, and REX1, but increased protein levels of FGF5 and cTnT (right panel). E MBD3 overexpression induced PKCi-derived mES differentiation and resulted in a loss of AP staining. Scale bar, 200 μm (upper panel). Overexpression of MBD3 reduced the total number of AP-positive colonies (left panel), decreased the percentage of undifferentiated and mixed colonies, and increased the percentage of differentiated colonies (right panel). Data were shown as mean ± SD (n = 3). The letters a and b indicated signiﬁcant differences among groups (P < 0.05).

Figure Legend Snippet: Fig. 4 Overexpression of MBD3-induced PKCi-derived mES differentiation. A qPCR showed that mRNA levels of MBD3 increased after PKCi- derived mES at passage 5 were transfected with FUW-MBD3, with the PKCi and FUW-M2rtTA plasmid groups used as controls. B MBD3 and β-actin protein levels were evaluated by Western blot after PKCi-derived mES were transfected with FUW-MBD3 (upper panel). Quantitative density analysis showed that protein levels of MBD3 increased after transfection with FUW-MBD3 (lower panel). C qPCR showed decreased mRNA levels of pluripotency genes NANOG and OCT4 and naïve-state marker REX1, but increased mRNA levels of primed-state marker FGF5 after MBD3 was overexpressed (left panel). MBD3 overexpression also increased mRNA levels of the endoderm marker CK8, mesoderm markers cTnT, BMP4, and DESMIN, and ectoderm markers PAX6 and SOX17 (right panel). D Western blot detection of NANOG, OCT4, REX1, FGF5, cTnT, and β-actin protein expression after MBD3 overexpression (left panel). Quantitative density analysis showed that MBD3 overexpression decreased protein levels of NANOG, OCT4, and REX1, but increased protein levels of FGF5 and cTnT (right panel). E MBD3 overexpression induced PKCi-derived mES differentiation and resulted in a loss of AP staining. Scale bar, 200 μm (upper panel). Overexpression of MBD3 reduced the total number of AP-positive colonies (left panel), decreased the percentage of undifferentiated and mixed colonies, and increased the percentage of differentiated colonies (right panel). Data were shown as mean ± SD (n = 3). The letters a and b indicated signiﬁcant differences among groups (P < 0.05).

Techniques Used: Over Expression, Derivative Assay, Transfection, Plasmid Preparation, Western Blot, Marker, Expressing, Staining

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Journal: Cell death discovery

Article Title: Nucleosome remodeling and deacetylation complex and MBD3 influence mouse embryonic stem cell naïve pluripotency under inhibition of protein kinase C.

doi: 10.1038/s41420-022-01131-0

Figure Lengend Snippet: Fig. 3 Knockdown of MBD3 promoted PKCi-derived mES self-renewal. A qPCR showed that mRNA levels of MBD3 decreased after PKCi- derived mES at passage 5 were transfected with shMBD3, with PKCi and shNC groups used as controls. B MBD3 and β-actin protein levels were evaluated by Western blot in PKCi-derived mES with MBD3 knockdown (upper panel). Quantitative density analysis showed that shMBD3 decreased protein levels of MBD3 (lower panel). C qPCR showed that compared with the control groups, shMBD3 increased the mRNA levels of pluripotency genes NANOG and OCT4 (left panel) but did not affect the mRNA levels of differentiation genes (right panel). D Western blot detection of NANOG, OCT4, and β-actin protein expression in PKCi-derived mES with MBD3 knockdown (left panel). Quantitative density analysis showed that shMBD3 increased protein levels of NANOG and OCT4 (right panel). E Knockdown of MBD3 did not affect the morphology or AP staining of PKCi-derived mES. Scale bar, 200 μm (upper panel). Knockdown of MBD3 did not affect the total number of AP- positive colonies (left panel) but increased the percentage of mixed colonies and decreased the percentage of differentiated colonies (right panel). Data were shown as mean ± SD (n = 3). The letters a and b indicated signiﬁcant differences among groups (P < 0.05).

Article Snippet: MBD3 overexpression in mES FUW-MBD3 (#52356) and control FUW-M2rtTA (#20342) were purchased from Addgene.

Techniques: Knockdown, Derivative Assay, Transfection, Western Blot, Control, Expressing, Staining

Journal: Cell death discovery

Article Title: Nucleosome remodeling and deacetylation complex and MBD3 influence mouse embryonic stem cell naïve pluripotency under inhibition of protein kinase C.

doi: 10.1038/s41420-022-01131-0

Figure Lengend Snippet: Fig. 4 Overexpression of MBD3-induced PKCi-derived mES differentiation. A qPCR showed that mRNA levels of MBD3 increased after PKCi- derived mES at passage 5 were transfected with FUW-MBD3, with the PKCi and FUW-M2rtTA plasmid groups used as controls. B MBD3 and β-actin protein levels were evaluated by Western blot after PKCi-derived mES were transfected with FUW-MBD3 (upper panel). Quantitative density analysis showed that protein levels of MBD3 increased after transfection with FUW-MBD3 (lower panel). C qPCR showed decreased mRNA levels of pluripotency genes NANOG and OCT4 and naïve-state marker REX1, but increased mRNA levels of primed-state marker FGF5 after MBD3 was overexpressed (left panel). MBD3 overexpression also increased mRNA levels of the endoderm marker CK8, mesoderm markers cTnT, BMP4, and DESMIN, and ectoderm markers PAX6 and SOX17 (right panel). D Western blot detection of NANOG, OCT4, REX1, FGF5, cTnT, and β-actin protein expression after MBD3 overexpression (left panel). Quantitative density analysis showed that MBD3 overexpression decreased protein levels of NANOG, OCT4, and REX1, but increased protein levels of FGF5 and cTnT (right panel). E MBD3 overexpression induced PKCi-derived mES differentiation and resulted in a loss of AP staining. Scale bar, 200 μm (upper panel). Overexpression of MBD3 reduced the total number of AP-positive colonies (left panel), decreased the percentage of undifferentiated and mixed colonies, and increased the percentage of differentiated colonies (right panel). Data were shown as mean ± SD (n = 3). The letters a and b indicated signiﬁcant differences among groups (P < 0.05).

Article Snippet: MBD3 overexpression in mES FUW-MBD3 (#52356) and control FUW-M2rtTA (#20342) were purchased from Addgene.

Techniques: Over Expression, Derivative Assay, Transfection, Plasmid Preparation, Western Blot, Marker, Expressing, Staining

Journal: American Journal of Translational Research

Article Title: Methyl-CpG-binding domain 3 (Mbd3) is an important regulator for apoptosis in mouse embryonic stem cells

doi:

Figure Lengend Snippet: Sequences of primers used for qPCR

Article Snippet: The Mbd3 overexpression plasmids, FUW-Mbd3 (#52356) and control FUW-M2rtTA (#20342), were purchased from Addgene.

Techniques:

Mbd3 overexpression induced cellular apoptosis. A. Overexpression of Mbd3 RNA levels was achieved by transfection with the FUW-Mbd3 plasmid in fifth-passage mES. The FUW-M2rtTA plasmid and the PKCi group were used as controls. B. Mbd3 protein expression was evaluated by Western blot in fifth-passage mES (upper panel). Quantitative density analysis showed that Mbd3 overexpression resulted in its increased protein levels compared with the PKCi control (lower panel). C. Mbd3 overexpression significantly reduced the total number of fifth-passage mES. D. CCK8 assay revealed that Mbd3 overexpression decreased mES cell viability. E. Apoptosis was evaluated by Annexin V staining in fifth-passage mES. The thick arrow indicates cell nuclei (blue), and the thin arrow indicates apoptotic cells (red). Scale bar =100 μm (left panel). The apoptosis rate was significantly increased with Mbd3 overexpression (right panel). F. Bax, Bcl-2, and β-actin protein levels were evaluated by Western blot in fifth-passage mES (left panel). Quantitative density analysis showed similar levels of Bax in all three groups (middle panel) and significantly reduced Bcl-2 levels with Mbd3 overexpression (right panel). G. Mbd3 overexpression increased the expression of Bax, Bim, Trail, Fasl, and caspase 3 RNA, but reduced the expression of Bcl-2 RNA, in fifth-passage mES. The data are represented as mean ± SD (n=3). *P<0.05, **P<0.01.

Journal: American Journal of Translational Research

Article Title: Methyl-CpG-binding domain 3 (Mbd3) is an important regulator for apoptosis in mouse embryonic stem cells

doi:

Figure Lengend Snippet: Mbd3 overexpression induced cellular apoptosis. A. Overexpression of Mbd3 RNA levels was achieved by transfection with the FUW-Mbd3 plasmid in fifth-passage mES. The FUW-M2rtTA plasmid and the PKCi group were used as controls. B. Mbd3 protein expression was evaluated by Western blot in fifth-passage mES (upper panel). Quantitative density analysis showed that Mbd3 overexpression resulted in its increased protein levels compared with the PKCi control (lower panel). C. Mbd3 overexpression significantly reduced the total number of fifth-passage mES. D. CCK8 assay revealed that Mbd3 overexpression decreased mES cell viability. E. Apoptosis was evaluated by Annexin V staining in fifth-passage mES. The thick arrow indicates cell nuclei (blue), and the thin arrow indicates apoptotic cells (red). Scale bar =100 μm (left panel). The apoptosis rate was significantly increased with Mbd3 overexpression (right panel). F. Bax, Bcl-2, and β-actin protein levels were evaluated by Western blot in fifth-passage mES (left panel). Quantitative density analysis showed similar levels of Bax in all three groups (middle panel) and significantly reduced Bcl-2 levels with Mbd3 overexpression (right panel). G. Mbd3 overexpression increased the expression of Bax, Bim, Trail, Fasl, and caspase 3 RNA, but reduced the expression of Bcl-2 RNA, in fifth-passage mES. The data are represented as mean ± SD (n=3). *P<0.05, **P<0.01.

Article Snippet: The Mbd3 overexpression plasmids, FUW-Mbd3 (#52356) and control FUW-M2rtTA (#20342), were purchased from Addgene.

Techniques: Over Expression, Transfection, Plasmid Preparation, Expressing, Western Blot, Control, CCK-8 Assay, Staining

Mbd3 knockdown partially reversed the cellular apoptosis induced by PKCi removal. A. Mbd3 RNA expression levels were increased in third-passage mES when PKCi was removed for 48 h from the culture medium, but they were only partially decreased with Mbd3 knockdown. B. Mbd3 protein levels were assessed by Western blot in third-passage mES (upper panel). Quantitative density analysis showed a significant increase in Mbd3 upon PKCi removal, and a partial reduction was observed with Mbd3 knockdown. C. The total number of third-passage mES was dramatically decreased upon PKCi removal and increased with Mbd3 knockdown, but it was still lower than that of the PKCi group. D. Cell viablility was improved by Mbd3 knockdown. E. Cellular apoptosis was assessed by Annexin V staining in third-passage mES. The thick arrow indicates cell nuclei (blue), and the thin arrow indicates apoptotic cells (red). Scale bar =100 μm (left panel). The apoptosis rate was significantly increased upon PKCi removal, but it was partially reduced with Mbd3 knockdown (right panel). F. Bax, Bcl-2, and β-actin levels were evaluated by Western blot in third-passage mES with Mbd3 knockdown (left panel). Quantitative density analysis showed increased levels of Bax after PKCi removal, but only partially reduced levels were observed with Mbd3 knockdown (middle panel). Bcl-2 expression was reduced upon PKCi removal, and similar levels were observed with Mbd3 knockdown (right panel). G. PKCi removal resulted in increased expression levels of Bax, Bim, Trail, Fasl, and caspase 3 RNA, with reduced Bcl-2 levels, in third-passage mES. Mbd3 knockdown resulted in partially reduced Bax, Bim, Trail, and caspase 3 RNA levels, unchanged Fasl levels, and increased Bcl-2 levels. The data are represented as mean ± SD (n=3). The letters a, b, c indicate significant differences among the groups (P<0.05).

Journal: American Journal of Translational Research

Article Title: Methyl-CpG-binding domain 3 (Mbd3) is an important regulator for apoptosis in mouse embryonic stem cells

doi:

Figure Lengend Snippet: Mbd3 knockdown partially reversed the cellular apoptosis induced by PKCi removal. A. Mbd3 RNA expression levels were increased in third-passage mES when PKCi was removed for 48 h from the culture medium, but they were only partially decreased with Mbd3 knockdown. B. Mbd3 protein levels were assessed by Western blot in third-passage mES (upper panel). Quantitative density analysis showed a significant increase in Mbd3 upon PKCi removal, and a partial reduction was observed with Mbd3 knockdown. C. The total number of third-passage mES was dramatically decreased upon PKCi removal and increased with Mbd3 knockdown, but it was still lower than that of the PKCi group. D. Cell viablility was improved by Mbd3 knockdown. E. Cellular apoptosis was assessed by Annexin V staining in third-passage mES. The thick arrow indicates cell nuclei (blue), and the thin arrow indicates apoptotic cells (red). Scale bar =100 μm (left panel). The apoptosis rate was significantly increased upon PKCi removal, but it was partially reduced with Mbd3 knockdown (right panel). F. Bax, Bcl-2, and β-actin levels were evaluated by Western blot in third-passage mES with Mbd3 knockdown (left panel). Quantitative density analysis showed increased levels of Bax after PKCi removal, but only partially reduced levels were observed with Mbd3 knockdown (middle panel). Bcl-2 expression was reduced upon PKCi removal, and similar levels were observed with Mbd3 knockdown (right panel). G. PKCi removal resulted in increased expression levels of Bax, Bim, Trail, Fasl, and caspase 3 RNA, with reduced Bcl-2 levels, in third-passage mES. Mbd3 knockdown resulted in partially reduced Bax, Bim, Trail, and caspase 3 RNA levels, unchanged Fasl levels, and increased Bcl-2 levels. The data are represented as mean ± SD (n=3). The letters a, b, c indicate significant differences among the groups (P<0.05).

Article Snippet: The Mbd3 overexpression plasmids, FUW-Mbd3 (#52356) and control FUW-M2rtTA (#20342), were purchased from Addgene.

Techniques: Knockdown, RNA Expression, Western Blot, Staining, Expressing

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1) Product Images from "Nucleosome remodeling and deacetylation complex and MBD3 influence mouse embryonic stem cell naïve pluripotency under inhibition of protein kinase C."

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