antiezh2 Search Results


p21  (Boster Bio)
94
Boster Bio p21
a Western blotting analysis showing that Neat1 knockdown enhanced <t>P21</t> protein expression but decreased Cdk4 protein expression in C2C12 cells. The relative protein levels of P21 and Cdk4 were quantified using ImageJ software. b , c ChIP-qPCR results revealed that the enrichments of Ezh2 ( b ) and H3k27me3 ( c ) at the P21 promoter were significantly decreased after Neat1 knockdown. d , e ChIP-qPCR results revealed that the enrichments of Ezh2 ( d ) and H3k27me3 ( e ) at the P21 promoter were significantly increased after Neat1 overexpression. f Co-transfection of Ezh2 siRNA fragment and Neat1 expression vector in C2C12 cells for 2 days. Immunofluorescence staining of P21 was performed, and P21 expression was quantified by ImageJ. The quantification of P21 immunofluorescence staining results showed that the overexpression of Neat1 inhibited P21 protein expression, but had no significant effect on P21 expression after co-transfection with Ezh2 siRNA fragment. g Neat1 siRNA fragment and Ezh2 expression vector were co-transfected into C2C12 cells and the cells were performed with EdU staining at 2 days after transfection. The percentage of EdU + cells was quantified. The quantification of EdU staining results showed that Neat1 knockdown inhibited myoblast proliferation. After co-transfected with Ezh2 expression vector, Neat1 knockdown can not inhibit myoblast proliferation. h Neat1 and P21 siRNA fragments were co-transfected into C2C12 cells and the cells were performed with EdU staining at 2 days after transfection. The quantification of EdU staining results showed that Neat1 knockdown significantly reduced the percentage of EdU + cells, but did not reduce the number of EdU + cells after co-transfection with P21 siRNA fragment. Protein levels were normalized to those of β-actin. All values represent the mean ± s.d. of three independent experiments. * p < 0.05, ** p < 0.01, N.S. indicates not significant
P21, supplied by Boster Bio, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/p21/product/Boster Bio
Average 94 stars, based on 1 article reviews
p21 - by Bioz Stars, 2026-03
94/100 stars
  Buy from Supplier
rabbit ezh2 antibody stj112944  (St Johns Laboratory)
90
St Johns Laboratory rabbit ezh2 antibody stj112944
a Western blotting analysis showing that Neat1 knockdown enhanced <t>P21</t> protein expression but decreased Cdk4 protein expression in C2C12 cells. The relative protein levels of P21 and Cdk4 were quantified using ImageJ software. b , c ChIP-qPCR results revealed that the enrichments of Ezh2 ( b ) and H3k27me3 ( c ) at the P21 promoter were significantly decreased after Neat1 knockdown. d , e ChIP-qPCR results revealed that the enrichments of Ezh2 ( d ) and H3k27me3 ( e ) at the P21 promoter were significantly increased after Neat1 overexpression. f Co-transfection of Ezh2 siRNA fragment and Neat1 expression vector in C2C12 cells for 2 days. Immunofluorescence staining of P21 was performed, and P21 expression was quantified by ImageJ. The quantification of P21 immunofluorescence staining results showed that the overexpression of Neat1 inhibited P21 protein expression, but had no significant effect on P21 expression after co-transfection with Ezh2 siRNA fragment. g Neat1 siRNA fragment and Ezh2 expression vector were co-transfected into C2C12 cells and the cells were performed with EdU staining at 2 days after transfection. The percentage of EdU + cells was quantified. The quantification of EdU staining results showed that Neat1 knockdown inhibited myoblast proliferation. After co-transfected with Ezh2 expression vector, Neat1 knockdown can not inhibit myoblast proliferation. h Neat1 and P21 siRNA fragments were co-transfected into C2C12 cells and the cells were performed with EdU staining at 2 days after transfection. The quantification of EdU staining results showed that Neat1 knockdown significantly reduced the percentage of EdU + cells, but did not reduce the number of EdU + cells after co-transfection with P21 siRNA fragment. Protein levels were normalized to those of β-actin. All values represent the mean ± s.d. of three independent experiments. * p < 0.05, ** p < 0.01, N.S. indicates not significant
Rabbit Ezh2 Antibody Stj112944, supplied by St Johns Laboratory, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit ezh2 antibody stj112944/product/St Johns Laboratory
Average 90 stars, based on 1 article reviews
rabbit ezh2 antibody stj112944 - by Bioz Stars, 2026-03
90/100 stars
  Buy from Supplier
secondary antibodies  (Boster Bio)
86
Boster Bio secondary antibodies
a Western blotting analysis showing that Neat1 knockdown enhanced <t>P21</t> protein expression but decreased Cdk4 protein expression in C2C12 cells. The relative protein levels of P21 and Cdk4 were quantified using ImageJ software. b , c ChIP-qPCR results revealed that the enrichments of Ezh2 ( b ) and H3k27me3 ( c ) at the P21 promoter were significantly decreased after Neat1 knockdown. d , e ChIP-qPCR results revealed that the enrichments of Ezh2 ( d ) and H3k27me3 ( e ) at the P21 promoter were significantly increased after Neat1 overexpression. f Co-transfection of Ezh2 siRNA fragment and Neat1 expression vector in C2C12 cells for 2 days. Immunofluorescence staining of P21 was performed, and P21 expression was quantified by ImageJ. The quantification of P21 immunofluorescence staining results showed that the overexpression of Neat1 inhibited P21 protein expression, but had no significant effect on P21 expression after co-transfection with Ezh2 siRNA fragment. g Neat1 siRNA fragment and Ezh2 expression vector were co-transfected into C2C12 cells and the cells were performed with EdU staining at 2 days after transfection. The percentage of EdU + cells was quantified. The quantification of EdU staining results showed that Neat1 knockdown inhibited myoblast proliferation. After co-transfected with Ezh2 expression vector, Neat1 knockdown can not inhibit myoblast proliferation. h Neat1 and P21 siRNA fragments were co-transfected into C2C12 cells and the cells were performed with EdU staining at 2 days after transfection. The quantification of EdU staining results showed that Neat1 knockdown significantly reduced the percentage of EdU + cells, but did not reduce the number of EdU + cells after co-transfection with P21 siRNA fragment. Protein levels were normalized to those of β-actin. All values represent the mean ± s.d. of three independent experiments. * p < 0.05, ** p < 0.01, N.S. indicates not significant
Secondary Antibodies, supplied by Boster Bio, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/secondary antibodies/product/Boster Bio
Average 86 stars, based on 1 article reviews
secondary antibodies - by Bioz Stars, 2026-03
86/100 stars
  Buy from Supplier
anti-ezh2  (Active Motif)
90
Active Motif anti-ezh2
a Western blotting analysis showing that Neat1 knockdown enhanced <t>P21</t> protein expression but decreased Cdk4 protein expression in C2C12 cells. The relative protein levels of P21 and Cdk4 were quantified using ImageJ software. b , c ChIP-qPCR results revealed that the enrichments of Ezh2 ( b ) and H3k27me3 ( c ) at the P21 promoter were significantly decreased after Neat1 knockdown. d , e ChIP-qPCR results revealed that the enrichments of Ezh2 ( d ) and H3k27me3 ( e ) at the P21 promoter were significantly increased after Neat1 overexpression. f Co-transfection of Ezh2 siRNA fragment and Neat1 expression vector in C2C12 cells for 2 days. Immunofluorescence staining of P21 was performed, and P21 expression was quantified by ImageJ. The quantification of P21 immunofluorescence staining results showed that the overexpression of Neat1 inhibited P21 protein expression, but had no significant effect on P21 expression after co-transfection with Ezh2 siRNA fragment. g Neat1 siRNA fragment and Ezh2 expression vector were co-transfected into C2C12 cells and the cells were performed with EdU staining at 2 days after transfection. The percentage of EdU + cells was quantified. The quantification of EdU staining results showed that Neat1 knockdown inhibited myoblast proliferation. After co-transfected with Ezh2 expression vector, Neat1 knockdown can not inhibit myoblast proliferation. h Neat1 and P21 siRNA fragments were co-transfected into C2C12 cells and the cells were performed with EdU staining at 2 days after transfection. The quantification of EdU staining results showed that Neat1 knockdown significantly reduced the percentage of EdU + cells, but did not reduce the number of EdU + cells after co-transfection with P21 siRNA fragment. Protein levels were normalized to those of β-actin. All values represent the mean ± s.d. of three independent experiments. * p < 0.05, ** p < 0.01, N.S. indicates not significant
Anti Ezh2, supplied by Active Motif, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti-ezh2/product/Active Motif
Average 90 stars, based on 1 article reviews
anti-ezh2 - by Bioz Stars, 2026-03
90/100 stars
  Buy from Supplier
anti-ezh2 antibody  (Becton Dickinson)
90
Becton Dickinson anti-ezh2 antibody
a Western blotting analysis showing that Neat1 knockdown enhanced <t>P21</t> protein expression but decreased Cdk4 protein expression in C2C12 cells. The relative protein levels of P21 and Cdk4 were quantified using ImageJ software. b , c ChIP-qPCR results revealed that the enrichments of Ezh2 ( b ) and H3k27me3 ( c ) at the P21 promoter were significantly decreased after Neat1 knockdown. d , e ChIP-qPCR results revealed that the enrichments of Ezh2 ( d ) and H3k27me3 ( e ) at the P21 promoter were significantly increased after Neat1 overexpression. f Co-transfection of Ezh2 siRNA fragment and Neat1 expression vector in C2C12 cells for 2 days. Immunofluorescence staining of P21 was performed, and P21 expression was quantified by ImageJ. The quantification of P21 immunofluorescence staining results showed that the overexpression of Neat1 inhibited P21 protein expression, but had no significant effect on P21 expression after co-transfection with Ezh2 siRNA fragment. g Neat1 siRNA fragment and Ezh2 expression vector were co-transfected into C2C12 cells and the cells were performed with EdU staining at 2 days after transfection. The percentage of EdU + cells was quantified. The quantification of EdU staining results showed that Neat1 knockdown inhibited myoblast proliferation. After co-transfected with Ezh2 expression vector, Neat1 knockdown can not inhibit myoblast proliferation. h Neat1 and P21 siRNA fragments were co-transfected into C2C12 cells and the cells were performed with EdU staining at 2 days after transfection. The quantification of EdU staining results showed that Neat1 knockdown significantly reduced the percentage of EdU + cells, but did not reduce the number of EdU + cells after co-transfection with P21 siRNA fragment. Protein levels were normalized to those of β-actin. All values represent the mean ± s.d. of three independent experiments. * p < 0.05, ** p < 0.01, N.S. indicates not significant
Anti Ezh2 Antibody, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti-ezh2 antibody/product/Becton Dickinson
Average 90 stars, based on 1 article reviews
anti-ezh2 antibody - by Bioz Stars, 2026-03
90/100 stars
  Buy from Supplier
ezh2  (Merck KGaA)
90
Merck KGaA ezh2
(A) MC3T3 cells were differentiated to mineralized osteoblasts for 9 days of culture (DIV) using Ascorbic Acid (AA) 50ug/mL. Nuclear extracts were collected at indicated the days and analyzed by western blot using specific antibodies against the indicated epigenetic regulators. TFIIB or RNA-polymerase II (RNAPII) was used to control for equal protein loading. (B-C and E-F) Binding of chromatin regulators to the Runx2 P1 promoter at the indicated differentiation days were analyzed by ChIP using antibodies against: (B) Wdr5, (C) Utx, (E) <t>Ezh2,</t> (F) Prmt5, and (G) Jarid1b. (D) Re-ChIP assay performed in chromatin samples obtained from differentiated cells (5 DIV) using first an antibody against Utx and subsequently an antibody against Wdr5. Results and statistical analyses are shown as described in figure legend 2. ***p<0.001, *p<0.05, ns = non-statistically significant differences.
Ezh2, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/ezh2/product/Merck KGaA
Average 90 stars, based on 1 article reviews
ezh2 - by Bioz Stars, 2026-03
90/100 stars
  Buy from Supplier
anti-ezh2  (Upstate Biotechnology Inc)
90
Upstate Biotechnology Inc anti-ezh2
(A) MC3T3 cells were differentiated to mineralized osteoblasts for 9 days of culture (DIV) using Ascorbic Acid (AA) 50ug/mL. Nuclear extracts were collected at indicated the days and analyzed by western blot using specific antibodies against the indicated epigenetic regulators. TFIIB or RNA-polymerase II (RNAPII) was used to control for equal protein loading. (B-C and E-F) Binding of chromatin regulators to the Runx2 P1 promoter at the indicated differentiation days were analyzed by ChIP using antibodies against: (B) Wdr5, (C) Utx, (E) <t>Ezh2,</t> (F) Prmt5, and (G) Jarid1b. (D) Re-ChIP assay performed in chromatin samples obtained from differentiated cells (5 DIV) using first an antibody against Utx and subsequently an antibody against Wdr5. Results and statistical analyses are shown as described in figure legend 2. ***p<0.001, *p<0.05, ns = non-statistically significant differences.
Anti Ezh2, supplied by Upstate Biotechnology Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti-ezh2/product/Upstate Biotechnology Inc
Average 90 stars, based on 1 article reviews
anti-ezh2 - by Bioz Stars, 2026-03
90/100 stars
  Buy from Supplier
anti-ezh2  (Rockland Immunochemicals)
90
Rockland Immunochemicals anti-ezh2
(A) MC3T3 cells were differentiated to mineralized osteoblasts for 9 days of culture (DIV) using Ascorbic Acid (AA) 50ug/mL. Nuclear extracts were collected at indicated the days and analyzed by western blot using specific antibodies against the indicated epigenetic regulators. TFIIB or RNA-polymerase II (RNAPII) was used to control for equal protein loading. (B-C and E-F) Binding of chromatin regulators to the Runx2 P1 promoter at the indicated differentiation days were analyzed by ChIP using antibodies against: (B) Wdr5, (C) Utx, (E) <t>Ezh2,</t> (F) Prmt5, and (G) Jarid1b. (D) Re-ChIP assay performed in chromatin samples obtained from differentiated cells (5 DIV) using first an antibody against Utx and subsequently an antibody against Wdr5. Results and statistical analyses are shown as described in figure legend 2. ***p<0.001, *p<0.05, ns = non-statistically significant differences.
Anti Ezh2, supplied by Rockland Immunochemicals, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti-ezh2/product/Rockland Immunochemicals
Average 90 stars, based on 1 article reviews
anti-ezh2 - by Bioz Stars, 2026-03
90/100 stars
  Buy from Supplier
anti-ezh2  (ABclonal Biotechnology)
90
ABclonal Biotechnology anti-ezh2
(A) MC3T3 cells were differentiated to mineralized osteoblasts for 9 days of culture (DIV) using Ascorbic Acid (AA) 50ug/mL. Nuclear extracts were collected at indicated the days and analyzed by western blot using specific antibodies against the indicated epigenetic regulators. TFIIB or RNA-polymerase II (RNAPII) was used to control for equal protein loading. (B-C and E-F) Binding of chromatin regulators to the Runx2 P1 promoter at the indicated differentiation days were analyzed by ChIP using antibodies against: (B) Wdr5, (C) Utx, (E) <t>Ezh2,</t> (F) Prmt5, and (G) Jarid1b. (D) Re-ChIP assay performed in chromatin samples obtained from differentiated cells (5 DIV) using first an antibody against Utx and subsequently an antibody against Wdr5. Results and statistical analyses are shown as described in figure legend 2. ***p<0.001, *p<0.05, ns = non-statistically significant differences.
Anti Ezh2, supplied by ABclonal Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti-ezh2/product/ABclonal Biotechnology
Average 90 stars, based on 1 article reviews
anti-ezh2 - by Bioz Stars, 2026-03
90/100 stars
  Buy from Supplier
ezh2  (Biocare Medical)
90
Biocare Medical ezh2
(A) MC3T3 cells were differentiated to mineralized osteoblasts for 9 days of culture (DIV) using Ascorbic Acid (AA) 50ug/mL. Nuclear extracts were collected at indicated the days and analyzed by western blot using specific antibodies against the indicated epigenetic regulators. TFIIB or RNA-polymerase II (RNAPII) was used to control for equal protein loading. (B-C and E-F) Binding of chromatin regulators to the Runx2 P1 promoter at the indicated differentiation days were analyzed by ChIP using antibodies against: (B) Wdr5, (C) Utx, (E) <t>Ezh2,</t> (F) Prmt5, and (G) Jarid1b. (D) Re-ChIP assay performed in chromatin samples obtained from differentiated cells (5 DIV) using first an antibody against Utx and subsequently an antibody against Wdr5. Results and statistical analyses are shown as described in figure legend 2. ***p<0.001, *p<0.05, ns = non-statistically significant differences.
Ezh2, supplied by Biocare Medical, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/ezh2/product/Biocare Medical
Average 90 stars, based on 1 article reviews
ezh2 - by Bioz Stars, 2026-03
90/100 stars
  Buy from Supplier
anti-ezh2  (MBL Life science)
90
MBL Life science anti-ezh2
(A) MC3T3 cells were differentiated to mineralized osteoblasts for 9 days of culture (DIV) using Ascorbic Acid (AA) 50ug/mL. Nuclear extracts were collected at indicated the days and analyzed by western blot using specific antibodies against the indicated epigenetic regulators. TFIIB or RNA-polymerase II (RNAPII) was used to control for equal protein loading. (B-C and E-F) Binding of chromatin regulators to the Runx2 P1 promoter at the indicated differentiation days were analyzed by ChIP using antibodies against: (B) Wdr5, (C) Utx, (E) <t>Ezh2,</t> (F) Prmt5, and (G) Jarid1b. (D) Re-ChIP assay performed in chromatin samples obtained from differentiated cells (5 DIV) using first an antibody against Utx and subsequently an antibody against Wdr5. Results and statistical analyses are shown as described in figure legend 2. ***p<0.001, *p<0.05, ns = non-statistically significant differences.
Anti Ezh2, supplied by MBL Life science, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti-ezh2/product/MBL Life science
Average 90 stars, based on 1 article reviews
anti-ezh2 - by Bioz Stars, 2026-03
90/100 stars
  Buy from Supplier
immunohistochemical stain for ezh2 11  (Cell Marque)
90
Cell Marque immunohistochemical stain for ezh2 11
(A) MC3T3 cells were differentiated to mineralized osteoblasts for 9 days of culture (DIV) using Ascorbic Acid (AA) 50ug/mL. Nuclear extracts were collected at indicated the days and analyzed by western blot using specific antibodies against the indicated epigenetic regulators. TFIIB or RNA-polymerase II (RNAPII) was used to control for equal protein loading. (B-C and E-F) Binding of chromatin regulators to the Runx2 P1 promoter at the indicated differentiation days were analyzed by ChIP using antibodies against: (B) Wdr5, (C) Utx, (E) <t>Ezh2,</t> (F) Prmt5, and (G) Jarid1b. (D) Re-ChIP assay performed in chromatin samples obtained from differentiated cells (5 DIV) using first an antibody against Utx and subsequently an antibody against Wdr5. Results and statistical analyses are shown as described in figure legend 2. ***p<0.001, *p<0.05, ns = non-statistically significant differences.
Immunohistochemical Stain For Ezh2 11, supplied by Cell Marque, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/immunohistochemical stain for ezh2 11/product/Cell Marque
Average 90 stars, based on 1 article reviews
immunohistochemical stain for ezh2 11 - by Bioz Stars, 2026-03
90/100 stars
  Buy from Supplier

Image Search Results


a Western blotting analysis showing that Neat1 knockdown enhanced P21 protein expression but decreased Cdk4 protein expression in C2C12 cells. The relative protein levels of P21 and Cdk4 were quantified using ImageJ software. b , c ChIP-qPCR results revealed that the enrichments of Ezh2 ( b ) and H3k27me3 ( c ) at the P21 promoter were significantly decreased after Neat1 knockdown. d , e ChIP-qPCR results revealed that the enrichments of Ezh2 ( d ) and H3k27me3 ( e ) at the P21 promoter were significantly increased after Neat1 overexpression. f Co-transfection of Ezh2 siRNA fragment and Neat1 expression vector in C2C12 cells for 2 days. Immunofluorescence staining of P21 was performed, and P21 expression was quantified by ImageJ. The quantification of P21 immunofluorescence staining results showed that the overexpression of Neat1 inhibited P21 protein expression, but had no significant effect on P21 expression after co-transfection with Ezh2 siRNA fragment. g Neat1 siRNA fragment and Ezh2 expression vector were co-transfected into C2C12 cells and the cells were performed with EdU staining at 2 days after transfection. The percentage of EdU + cells was quantified. The quantification of EdU staining results showed that Neat1 knockdown inhibited myoblast proliferation. After co-transfected with Ezh2 expression vector, Neat1 knockdown can not inhibit myoblast proliferation. h Neat1 and P21 siRNA fragments were co-transfected into C2C12 cells and the cells were performed with EdU staining at 2 days after transfection. The quantification of EdU staining results showed that Neat1 knockdown significantly reduced the percentage of EdU + cells, but did not reduce the number of EdU + cells after co-transfection with P21 siRNA fragment. Protein levels were normalized to those of β-actin. All values represent the mean ± s.d. of three independent experiments. * p < 0.05, ** p < 0.01, N.S. indicates not significant

Journal: Cell Death & Disease

Article Title: Long noncoding RNA Neat1 modulates myogenesis by recruiting Ezh2

doi: 10.1038/s41419-019-1742-7

Figure Lengend Snippet: a Western blotting analysis showing that Neat1 knockdown enhanced P21 protein expression but decreased Cdk4 protein expression in C2C12 cells. The relative protein levels of P21 and Cdk4 were quantified using ImageJ software. b , c ChIP-qPCR results revealed that the enrichments of Ezh2 ( b ) and H3k27me3 ( c ) at the P21 promoter were significantly decreased after Neat1 knockdown. d , e ChIP-qPCR results revealed that the enrichments of Ezh2 ( d ) and H3k27me3 ( e ) at the P21 promoter were significantly increased after Neat1 overexpression. f Co-transfection of Ezh2 siRNA fragment and Neat1 expression vector in C2C12 cells for 2 days. Immunofluorescence staining of P21 was performed, and P21 expression was quantified by ImageJ. The quantification of P21 immunofluorescence staining results showed that the overexpression of Neat1 inhibited P21 protein expression, but had no significant effect on P21 expression after co-transfection with Ezh2 siRNA fragment. g Neat1 siRNA fragment and Ezh2 expression vector were co-transfected into C2C12 cells and the cells were performed with EdU staining at 2 days after transfection. The percentage of EdU + cells was quantified. The quantification of EdU staining results showed that Neat1 knockdown inhibited myoblast proliferation. After co-transfected with Ezh2 expression vector, Neat1 knockdown can not inhibit myoblast proliferation. h Neat1 and P21 siRNA fragments were co-transfected into C2C12 cells and the cells were performed with EdU staining at 2 days after transfection. The quantification of EdU staining results showed that Neat1 knockdown significantly reduced the percentage of EdU + cells, but did not reduce the number of EdU + cells after co-transfection with P21 siRNA fragment. Protein levels were normalized to those of β-actin. All values represent the mean ± s.d. of three independent experiments. * p < 0.05, ** p < 0.01, N.S. indicates not significant

Article Snippet: The antibodies and their dilutions were shown as following: Myod (Santa Cruz Biotechnology, USA; sc-760; 1:1000), Myog (Santa Cruz Biotechnology, USA; sc-12732; 1:200), Myhc (Santa Cruz Biotechnology, USA; sc-376157; 1:3000), α-actin (Proteintech, China; 23660-1-AP; 1:1000), Tnni2 (Abcam, UK; ab184554; 1:1000), P21 (BOSTER, China; BM4382; 1:200), Ezh2 (Cell Signaling Technology, USA; 5246; 1:1000), Pcna (Servicebio, China; GB11010; 1:500), Ki67 (Abcam, UK; ab16667; 1:1000), β-actin (Santa Cruz Biotechnology, USA; sc-4777; 1:1000), Gapdh (BOSTER, China; BM3876; 1:200), Pax7 (Developmental Studies Hybridoma Bank; USA; 1:1000), eMyhc (Developmental Studies Hybridoma Bank, USA; BF-G6; 1:1000).

Techniques: Western Blot, Knockdown, Expressing, Software, ChIP-qPCR, Over Expression, Cotransfection, Plasmid Preparation, Immunofluorescence, Staining, Transfection

In proliferating myoblasts, Neat1 guides Ezh2 to the P21 promoter and inhibits P21 expression, leading to the promotion of myoblast proliferation. Upon differentiation, Neat1 recruits Ezh2 to inhibit the expression of muscle-specific genes, such as Myog , Myh4 , and Tnni2 , and suppresses myogenic differentiation

Journal: Cell Death & Disease

Article Title: Long noncoding RNA Neat1 modulates myogenesis by recruiting Ezh2

doi: 10.1038/s41419-019-1742-7

Figure Lengend Snippet: In proliferating myoblasts, Neat1 guides Ezh2 to the P21 promoter and inhibits P21 expression, leading to the promotion of myoblast proliferation. Upon differentiation, Neat1 recruits Ezh2 to inhibit the expression of muscle-specific genes, such as Myog , Myh4 , and Tnni2 , and suppresses myogenic differentiation

Article Snippet: The antibodies and their dilutions were shown as following: Myod (Santa Cruz Biotechnology, USA; sc-760; 1:1000), Myog (Santa Cruz Biotechnology, USA; sc-12732; 1:200), Myhc (Santa Cruz Biotechnology, USA; sc-376157; 1:3000), α-actin (Proteintech, China; 23660-1-AP; 1:1000), Tnni2 (Abcam, UK; ab184554; 1:1000), P21 (BOSTER, China; BM4382; 1:200), Ezh2 (Cell Signaling Technology, USA; 5246; 1:1000), Pcna (Servicebio, China; GB11010; 1:500), Ki67 (Abcam, UK; ab16667; 1:1000), β-actin (Santa Cruz Biotechnology, USA; sc-4777; 1:1000), Gapdh (BOSTER, China; BM3876; 1:200), Pax7 (Developmental Studies Hybridoma Bank; USA; 1:1000), eMyhc (Developmental Studies Hybridoma Bank, USA; BF-G6; 1:1000).

Techniques: Expressing

(A) MC3T3 cells were differentiated to mineralized osteoblasts for 9 days of culture (DIV) using Ascorbic Acid (AA) 50ug/mL. Nuclear extracts were collected at indicated the days and analyzed by western blot using specific antibodies against the indicated epigenetic regulators. TFIIB or RNA-polymerase II (RNAPII) was used to control for equal protein loading. (B-C and E-F) Binding of chromatin regulators to the Runx2 P1 promoter at the indicated differentiation days were analyzed by ChIP using antibodies against: (B) Wdr5, (C) Utx, (E) Ezh2, (F) Prmt5, and (G) Jarid1b. (D) Re-ChIP assay performed in chromatin samples obtained from differentiated cells (5 DIV) using first an antibody against Utx and subsequently an antibody against Wdr5. Results and statistical analyses are shown as described in figure legend 2. ***p<0.001, *p<0.05, ns = non-statistically significant differences.

Journal: Journal of cellular physiology

Article Title: Mll-COMPASS complexes mediate H3K4me3 enrichment and transcription of the osteoblast master gene Runx2/p57 in osteoblasts

doi: 10.1002/jcp.27355

Figure Lengend Snippet: (A) MC3T3 cells were differentiated to mineralized osteoblasts for 9 days of culture (DIV) using Ascorbic Acid (AA) 50ug/mL. Nuclear extracts were collected at indicated the days and analyzed by western blot using specific antibodies against the indicated epigenetic regulators. TFIIB or RNA-polymerase II (RNAPII) was used to control for equal protein loading. (B-C and E-F) Binding of chromatin regulators to the Runx2 P1 promoter at the indicated differentiation days were analyzed by ChIP using antibodies against: (B) Wdr5, (C) Utx, (E) Ezh2, (F) Prmt5, and (G) Jarid1b. (D) Re-ChIP assay performed in chromatin samples obtained from differentiated cells (5 DIV) using first an antibody against Utx and subsequently an antibody against Wdr5. Results and statistical analyses are shown as described in figure legend 2. ***p<0.001, *p<0.05, ns = non-statistically significant differences.

Article Snippet: The following antibodies were used in ChIP assays: Wdr5 (ab56919, Abcam), Jarid1b/Kdm4b (ab50958, Abcam), Ezh2 (07–689, Merck Millipore, Danvers, MA, USA), Utx/Kdm6a (ab91231, Abcam), Prmt5/Jbp1 (611539, BD Biosciences), H3K27me3 (07–449, Merck Millipore), H3K4me1 (ab8895, Abcam), H3K4me3 (ab8580, Abcam), H4R3me2S (ab5823, Abcam), H3Ac (06–599, Merck Millipore), Cgbp (SC-25391, Santa Cruz Biotechnology), Menin (A300–105A, Bethyl Laboratories, Montgomery, TX, USA), Set1A (A300–290A, Bethyl Laboratories), Set1B (SC-248563, Santa Cruz Biotechnology), Mll1 (39829, Active Motif), Mll2 (SC-292359, Santa Cruz Biotechnology), Mll3 (ab71200, Abcam), Mll4 (ab60053, Abcam).

Techniques: Western Blot, Binding Assay

(A-B and D-E) Differentiating MC3T3 cells (3DIV) were infected with lentiviral particles containing shRNAs against the chromatin modifiers: (A) Wdr5, (B) Utx, (D) Ezh2 and (E) Prmt5. Knockdown efficiencies were confirmed by RT-qPCR and western blot (A-D, left panels) analyses, 48 h post-infection (5 DIV). TFIIB or RNAPII proteins were used to control for equal protein loading. (C) Effect of knocking down Wdr5 (left) and Utx (right) expression on Bglap gene transcription. mRNA expression values were normalized against Gapdh mRNA levels. Statistical analyses were assessed with respect to the mRNA levels obtained in cells infected with an empty vector (EV). *p<0.05, ***p<0.001, ns = non-statistically significant differences.

Journal: Journal of cellular physiology

Article Title: Mll-COMPASS complexes mediate H3K4me3 enrichment and transcription of the osteoblast master gene Runx2/p57 in osteoblasts

doi: 10.1002/jcp.27355

Figure Lengend Snippet: (A-B and D-E) Differentiating MC3T3 cells (3DIV) were infected with lentiviral particles containing shRNAs against the chromatin modifiers: (A) Wdr5, (B) Utx, (D) Ezh2 and (E) Prmt5. Knockdown efficiencies were confirmed by RT-qPCR and western blot (A-D, left panels) analyses, 48 h post-infection (5 DIV). TFIIB or RNAPII proteins were used to control for equal protein loading. (C) Effect of knocking down Wdr5 (left) and Utx (right) expression on Bglap gene transcription. mRNA expression values were normalized against Gapdh mRNA levels. Statistical analyses were assessed with respect to the mRNA levels obtained in cells infected with an empty vector (EV). *p<0.05, ***p<0.001, ns = non-statistically significant differences.

Article Snippet: The following antibodies were used in ChIP assays: Wdr5 (ab56919, Abcam), Jarid1b/Kdm4b (ab50958, Abcam), Ezh2 (07–689, Merck Millipore, Danvers, MA, USA), Utx/Kdm6a (ab91231, Abcam), Prmt5/Jbp1 (611539, BD Biosciences), H3K27me3 (07–449, Merck Millipore), H3K4me1 (ab8895, Abcam), H3K4me3 (ab8580, Abcam), H4R3me2S (ab5823, Abcam), H3Ac (06–599, Merck Millipore), Cgbp (SC-25391, Santa Cruz Biotechnology), Menin (A300–105A, Bethyl Laboratories, Montgomery, TX, USA), Set1A (A300–290A, Bethyl Laboratories), Set1B (SC-248563, Santa Cruz Biotechnology), Mll1 (39829, Active Motif), Mll2 (SC-292359, Santa Cruz Biotechnology), Mll3 (ab71200, Abcam), Mll4 (ab60053, Abcam).

Techniques: Infection, Quantitative RT-PCR, Western Blot, Expressing, Plasmid Preparation