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Novus Biologicals rabbit polyclonal anti chd8
Rabbit Polyclonal Anti Chd8, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 91/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/chd8+rabbit+polyclonal+antibody/CHD8+Antibody/pmc12204788-953-0-4
Average 91 stars, based on 6 article reviews

rabbit polyclonal anti chd8 - by Bioz Stars, 2026-10

91/100 stars

Images

Transfection:

Article Title: Widespread impact of nucleosome remodelers on transcription at cis-regulatory elements
Article Snippet: The following antibodies were used in CUT&RUN experiments performed on 100,000 lightly crosslinked wildtype ES cells using a low salt elution method for fragment release: SRCAP (Kerafast ESL103,1:50) SMARCAL1 (Invitrogen PA5-28980, lot YF3945097B, 1:50), and IgG (Sigma-Aldrich, 06-371,1:250). .. The following antibodies were used in low salt elution uncrosslinked CUT&RUN experiments on 100,000 cells transfected by esiRNAs: H3K27ac (abcam ab4729, lot GR3416784-1, 1:100), H2A.Z (abcam ab4174, lot GR3198864-1, 1:100), NFYA (Santa Cruz SC-17753, lot D2522, 1:50), SRCAP (Kerafast ESL103,1:50) SMARCAL1 (Invitrogen PA5-28980, lot YF3945097B, 1:50), CHD8 (NOVUS NB100-60418 lot A4), and IgG (Sigma-Aldrich, 06-371,1:250). ..

other:

Article Title: Widespread impact of nucleosome remodelers on transcription at cis-regulatory elements
Article Snippet: Rabbit polyclonal anti-CHD8 , NOVUS , nb10060418; RRID:AB_905325.

Image Search Results

Journal: iScience

Article Title: A CHD8-TRRAP axis facilitates MYC and E2F target gene regulation in human neural stem cells

doi: 10.1016/j.isci.2025.111978

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal Anti-CHD8 , Bethyl , Cat# A301-224A; RRID: AB_890578.

Techniques: Virus, Recombinant, Membrane, Protease Inhibitor, SYBR Green Assay, Staining, cDNA Synthesis, Mass Spectrometry, shRNA, Sequencing, Control, Plasmid Preparation, Software, Sonication

(A) Genetic strategy targeting exon 4 of the Chd8 gene. The exon was flanked with loxP sites to excise the loci, resulting in a frameshift mutation that disrupts the production of the CHD8 protein after Cre excision. (B and C) Representative images of CHD8 in the CA1 regions of adult mouse brains. CHD8 is expressed in neurons and astrocytes in controls (B). After tamoxifen administration, Chd8 fx/fx : CAGGS-CreER +/ − mouse brains showed non-detectable levels of CHD8 protein (C). In both (B) and (C), arrows point to astrocytes expressing CHD8 and its knockout thereafter. (D) Schematic diagram for tamoxifen-induced Chd8 cKO, the stab-wound injury model, and the analysis of reactive gliosis. (E and F) Response of GFAP + astrocytes and Iba1 + microglia after stab-wound injury. Control mice (E) exhibit astrocytic and microglial response as expected, while global Chd8 cKO mice (F) show reduced staining for both GFAP + and Iba1 + along the needle track. (G and H) High-magnification images from the injury site from the corresponding genotypes. Note the reduction in cell body size, process elongation, and polarity in astrocytes from global Chd8 cKO mice (H). (I and J) Decreased area occupied by GFAP + astrocytes (I) and Iba1 + microglia (J) in global Chd8 cKO mice compared to controls. In (B) and (C), scale bars indicate 20 μm; in (G) and (H), scale bars indicate 50 μm; in (E) and (F), scale bars indicate 500 μm. The dashed rectangle indicates the ROIs that were quantified. In (E)–(H), the dashed lines indicate the needle track of the injury. In (I) and (J), data points illustrate the quantified area from the six brain slices most proximal to the injury epicenter. Data are normalized to the means of the ipsilateral site in control mice. **** p < 0.0001, ns, not significant; statistical analysis was performed with two-way ANOVA; on the violin plots, dashed lines indicate the 25%, mean, and 75% percentile, respectively, from bottom to top; n = 4 mice per genotype.

Journal: Cell reports

Article Title: Autism-associated CHD8 controls reactive gliosis and neuroinflammation via remodeling chromatin in astrocytes

doi: 10.1016/j.celrep.2024.114637

Figure Lengend Snippet: (A) Genetic strategy targeting exon 4 of the Chd8 gene. The exon was flanked with loxP sites to excise the loci, resulting in a frameshift mutation that disrupts the production of the CHD8 protein after Cre excision. (B and C) Representative images of CHD8 in the CA1 regions of adult mouse brains. CHD8 is expressed in neurons and astrocytes in controls (B). After tamoxifen administration, Chd8 fx/fx : CAGGS-CreER +/ − mouse brains showed non-detectable levels of CHD8 protein (C). In both (B) and (C), arrows point to astrocytes expressing CHD8 and its knockout thereafter. (D) Schematic diagram for tamoxifen-induced Chd8 cKO, the stab-wound injury model, and the analysis of reactive gliosis. (E and F) Response of GFAP + astrocytes and Iba1 + microglia after stab-wound injury. Control mice (E) exhibit astrocytic and microglial response as expected, while global Chd8 cKO mice (F) show reduced staining for both GFAP + and Iba1 + along the needle track. (G and H) High-magnification images from the injury site from the corresponding genotypes. Note the reduction in cell body size, process elongation, and polarity in astrocytes from global Chd8 cKO mice (H). (I and J) Decreased area occupied by GFAP + astrocytes (I) and Iba1 + microglia (J) in global Chd8 cKO mice compared to controls. In (B) and (C), scale bars indicate 20 μm; in (G) and (H), scale bars indicate 50 μm; in (E) and (F), scale bars indicate 500 μm. The dashed rectangle indicates the ROIs that were quantified. In (E)–(H), the dashed lines indicate the needle track of the injury. In (I) and (J), data points illustrate the quantified area from the six brain slices most proximal to the injury epicenter. Data are normalized to the means of the ipsilateral site in control mice. **** p < 0.0001, ns, not significant; statistical analysis was performed with two-way ANOVA; on the violin plots, dashed lines indicate the 25%, mean, and 75% percentile, respectively, from bottom to top; n = 4 mice per genotype.

Article Snippet: CHD8 Rabbit polyclonal 1 to 1000 , Novus Biologicals , NB 100-60418.

Techniques: Mutagenesis, Expressing, Knock-Out, Control, Staining

(A) Strategy for tamoxifen-induced, astrocyte-specific Chd8 cKO utilizing the Aldh1l1-CreERT2 line. Mice were crossed with the Ai14- tdTomato reporter line to visualize recombined cells. (B and C) Astrocyte-specific Chd8- cKO mice show non-detectable CHD8 protein expression in cortical slices. Controls are mice without the Chd8 floxed alleles but expressing Aldh1l1-CreERT2 to turn on the expression of tdTomato reporter. Arrows indicate CHD8 expression in tdTomato + astrocytes in control but not astrocyte Chd8 cKO. (D and E) GFAP and Iba1 staining of astrocytes and microglia, respectively, after stab-wound injury in control (D) and astrocyte cKO mice (E). (F and G) High-magnification images from the injury sites from the corresponding genotypes showing reduction in cell body size, process elongation, and polarity in astrocytes and reduced microglia numbers in astrocyte cKO mice. (H and I) Decreased area occupied by GFAP + astrocytes and their numbers in astrocyte cKO mice compared to controls. (J and K) Decreased area occupied by Iba1 + microglia and their numbers in astrocyte cKO mice compared to controls. In (H)–(K), data points represent the quantified area from the six brain slices most proximal to the injury epicenter. Data are normalized to the means of the ipsilateral site in control mice. In (B) and (C), scale bars indicate 20 μm; in (F) and (G), scale bars indicate 50 μm; in (D) and (E), scale bars indicate 500 μm. The dashed rectangle indicates the ROIs that were quantified. In (D)–(G), the dashed lines in merged images indicate the needle track of the injury. In (H)–(K), *** p < 0.001; **** p < 0.0001; ns, not significant; statistical comparisons were analyzed with two-way ANOVA; on the violin plots, dashed lines indicate the 25%, mean, and 75% percentile, respectively, from bottom to top; n = 6 mice per genotype.

Journal: Cell reports

Article Title: Autism-associated CHD8 controls reactive gliosis and neuroinflammation via remodeling chromatin in astrocytes

doi: 10.1016/j.celrep.2024.114637

Figure Lengend Snippet: (A) Strategy for tamoxifen-induced, astrocyte-specific Chd8 cKO utilizing the Aldh1l1-CreERT2 line. Mice were crossed with the Ai14- tdTomato reporter line to visualize recombined cells. (B and C) Astrocyte-specific Chd8- cKO mice show non-detectable CHD8 protein expression in cortical slices. Controls are mice without the Chd8 floxed alleles but expressing Aldh1l1-CreERT2 to turn on the expression of tdTomato reporter. Arrows indicate CHD8 expression in tdTomato + astrocytes in control but not astrocyte Chd8 cKO. (D and E) GFAP and Iba1 staining of astrocytes and microglia, respectively, after stab-wound injury in control (D) and astrocyte cKO mice (E). (F and G) High-magnification images from the injury sites from the corresponding genotypes showing reduction in cell body size, process elongation, and polarity in astrocytes and reduced microglia numbers in astrocyte cKO mice. (H and I) Decreased area occupied by GFAP + astrocytes and their numbers in astrocyte cKO mice compared to controls. (J and K) Decreased area occupied by Iba1 + microglia and their numbers in astrocyte cKO mice compared to controls. In (H)–(K), data points represent the quantified area from the six brain slices most proximal to the injury epicenter. Data are normalized to the means of the ipsilateral site in control mice. In (B) and (C), scale bars indicate 20 μm; in (F) and (G), scale bars indicate 50 μm; in (D) and (E), scale bars indicate 500 μm. The dashed rectangle indicates the ROIs that were quantified. In (D)–(G), the dashed lines in merged images indicate the needle track of the injury. In (H)–(K), *** p < 0.001; **** p < 0.0001; ns, not significant; statistical comparisons were analyzed with two-way ANOVA; on the violin plots, dashed lines indicate the 25%, mean, and 75% percentile, respectively, from bottom to top; n = 6 mice per genotype.

Article Snippet: CHD8 Rabbit polyclonal 1 to 1000 , Novus Biologicals , NB 100-60418.

Techniques: Expressing, Control, Staining

(A) Heatmap of DEGs identified through RNA-seq between control and astrocyte Chd8 cKO mice after LPS treatment ( n = 5 mice per genotype). A total of 109 DEGs were identified, 76 of which were upregulated and 33 were downregulated (FDR < 0.05). (B) Volcano plot depicting the distribution of upregulated and downregulated genes, relative to their quantified fold change and their corresponding p values. The threshold was set at p (adjusted) < 0.05. (C) Venn diagram of the detected DEGs, depicting a subset of DEGs that correspond to genes whose expression is specific to astrocytes, neurons, and microglia. Of those, many DEGs (41) were determined to be astrocyte specific, while fewer were deemed to be specific in neurons (4) and microglia (1). (D) Bar plot showing the fold enrichment of the detected DEGs in our dataset, indicating significant enrichment of DEGs for astrocyte marker genes. Notably, no enrichment was detected when comparing neuronal or microglial genes to the cell-type markers from previous studies, as cited. *** p < 0.001; **** p < 0.0001. (E) GO terms analysis reveals changes associated with many cellular processes, including lipid and metabolic pathways in astrocyte cKO mice in response to LPS stimulation. (F) Heatmap of the top 20 DEGs identified through RNA-seq. Of these, 13 were upregulated and 7 were downregulated in astrocyte cKO mice treated with LPS vs. control mice treated with LPS. (G–O) qPCR analysis of Gstt3 (G), Acsl3 (H), Etnppl (I), Phykpl (J), Gjb6 (K), Slc9a8 (L), Agt (M), Tnfrsf25 (N), and Lcat (O) mRNA confirms the altered expression shown in (F) ( n = 4 mice per group). (P) Representative western blotting of TNFRSF25 from cortices of control and astrocyte cKO mice after LPS administration, showing reduced TNFRSF25 protein in the cortex of astrocyte cKO mice ( n = 7 mice per group). (Q) Representative western blotting of LCAT from cortices of control and astrocyte cKO mice after LPS administration showing reduced LCAT protein in astrocyte cKO mice ( n = 7). (R) Representative images of TNFRSF25 staining in the cortex of control and astrocyte cKO mice after LPS administration. Signal intensity quantification of TNFRSF25 staining ( n = 4 mice per genotype). * p < 0.05; scale bars indicate 200 μm. Data points are normalized to the mean of the control group. Error bars depict the SEM. Statistical comparisons were performed with the one-tailed Welch’s t test.

Journal: Cell reports

Article Title: Autism-associated CHD8 controls reactive gliosis and neuroinflammation via remodeling chromatin in astrocytes

doi: 10.1016/j.celrep.2024.114637

Figure Lengend Snippet: (A) Heatmap of DEGs identified through RNA-seq between control and astrocyte Chd8 cKO mice after LPS treatment ( n = 5 mice per genotype). A total of 109 DEGs were identified, 76 of which were upregulated and 33 were downregulated (FDR < 0.05). (B) Volcano plot depicting the distribution of upregulated and downregulated genes, relative to their quantified fold change and their corresponding p values. The threshold was set at p (adjusted) < 0.05. (C) Venn diagram of the detected DEGs, depicting a subset of DEGs that correspond to genes whose expression is specific to astrocytes, neurons, and microglia. Of those, many DEGs (41) were determined to be astrocyte specific, while fewer were deemed to be specific in neurons (4) and microglia (1). (D) Bar plot showing the fold enrichment of the detected DEGs in our dataset, indicating significant enrichment of DEGs for astrocyte marker genes. Notably, no enrichment was detected when comparing neuronal or microglial genes to the cell-type markers from previous studies, as cited. *** p < 0.001; **** p < 0.0001. (E) GO terms analysis reveals changes associated with many cellular processes, including lipid and metabolic pathways in astrocyte cKO mice in response to LPS stimulation. (F) Heatmap of the top 20 DEGs identified through RNA-seq. Of these, 13 were upregulated and 7 were downregulated in astrocyte cKO mice treated with LPS vs. control mice treated with LPS. (G–O) qPCR analysis of Gstt3 (G), Acsl3 (H), Etnppl (I), Phykpl (J), Gjb6 (K), Slc9a8 (L), Agt (M), Tnfrsf25 (N), and Lcat (O) mRNA confirms the altered expression shown in (F) ( n = 4 mice per group). (P) Representative western blotting of TNFRSF25 from cortices of control and astrocyte cKO mice after LPS administration, showing reduced TNFRSF25 protein in the cortex of astrocyte cKO mice ( n = 7 mice per group). (Q) Representative western blotting of LCAT from cortices of control and astrocyte cKO mice after LPS administration showing reduced LCAT protein in astrocyte cKO mice ( n = 7). (R) Representative images of TNFRSF25 staining in the cortex of control and astrocyte cKO mice after LPS administration. Signal intensity quantification of TNFRSF25 staining ( n = 4 mice per genotype). * p < 0.05; scale bars indicate 200 μm. Data points are normalized to the mean of the control group. Error bars depict the SEM. Statistical comparisons were performed with the one-tailed Welch’s t test.

Article Snippet: CHD8 Rabbit polyclonal 1 to 1000 , Novus Biologicals , NB 100-60418.

Techniques: RNA Sequencing, Control, Expressing, Marker, Western Blot, Staining, One-tailed Test

(A) Schematic diagram illustrating the elements required for the designed AAV for astrocyte-specific Chd8 editing in vivo via CRISPR-SaCas9. (B) Diagram for the simultaneous AAV injection and stab-wound injury with analysis of reactive gliosis performed at 7 days post-injection. (C) Representative images near the needle track from control mice (Scramble-AAV injected). CHD8 is detectable in astrocytes (SOX9 + ) expressing SaCas9 (HA tag + ). (D) Representative images near the needle track from mice injected with the Chd8 -cKO AAVs. CHD8 is undetectable in the majority of HA + and SOX9 + astrocytes (white arrows), while fewer HA + and SOX9 + astrocytes still show CHD8 expression (yellow arrows). (E and F) GFAP and Iba1 staining of astrocytes and microglia, respectively, after stab-wound injury and AAV injection in the Scramble-AAV (E) and Chd8 -cKO-AAV groups (F). (G) Decreased area occupied by GFAP + astrocytes in the Chd8 -cKO-AAV mice. (H) Quantification of the area occupied by Iba1 + microglia between control and Chd8 -cKO-AAV mice. In (C) and (D), scale bars indicate 20 μm; in (E) and (F), scale bars indicate 500 μm. The dashed rectangle indicates the ROIs that were quantified in (G) and (H). The dashed lines in merged images indicate the needle track of the injury. In (G) and (H), statistical comparisons were performed with two-way ANOVA; ** p < 0.01. Data points indicate n = 3 mice per group. Data are normalized to the means of the ipsilateral site in the control group.

Journal: Cell reports

Article Title: Autism-associated CHD8 controls reactive gliosis and neuroinflammation via remodeling chromatin in astrocytes

doi: 10.1016/j.celrep.2024.114637

Figure Lengend Snippet: (A) Schematic diagram illustrating the elements required for the designed AAV for astrocyte-specific Chd8 editing in vivo via CRISPR-SaCas9. (B) Diagram for the simultaneous AAV injection and stab-wound injury with analysis of reactive gliosis performed at 7 days post-injection. (C) Representative images near the needle track from control mice (Scramble-AAV injected). CHD8 is detectable in astrocytes (SOX9 + ) expressing SaCas9 (HA tag + ). (D) Representative images near the needle track from mice injected with the Chd8 -cKO AAVs. CHD8 is undetectable in the majority of HA + and SOX9 + astrocytes (white arrows), while fewer HA + and SOX9 + astrocytes still show CHD8 expression (yellow arrows). (E and F) GFAP and Iba1 staining of astrocytes and microglia, respectively, after stab-wound injury and AAV injection in the Scramble-AAV (E) and Chd8 -cKO-AAV groups (F). (G) Decreased area occupied by GFAP + astrocytes in the Chd8 -cKO-AAV mice. (H) Quantification of the area occupied by Iba1 + microglia between control and Chd8 -cKO-AAV mice. In (C) and (D), scale bars indicate 20 μm; in (E) and (F), scale bars indicate 500 μm. The dashed rectangle indicates the ROIs that were quantified in (G) and (H). The dashed lines in merged images indicate the needle track of the injury. In (G) and (H), statistical comparisons were performed with two-way ANOVA; ** p < 0.01. Data points indicate n = 3 mice per group. Data are normalized to the means of the ipsilateral site in the control group.

Article Snippet: CHD8 Rabbit polyclonal 1 to 1000 , Novus Biologicals , NB 100-60418.

Techniques: In Vivo, CRISPR, Injection, Control, Expressing, Staining

Figure 1. Reduced reactive gliosis in the global Chd8-cKO mice in the stab-wound injury model (A) Genetic strategy targeting exon 4 of the Chd8 gene. The exon was ﬂanked with loxP sites to excise the loci, resulting in a frameshift mutation that disrupts the production of the CHD8 protein after Cre excision.

Journal: Cell reports

Article Title: Autism-associated CHD8 controls reactive gliosis and neuroinflammation via remodeling chromatin in astrocytes.

doi: 10.1016/j.celrep.2024.114637

Figure Lengend Snippet: Figure 1. Reduced reactive gliosis in the global Chd8-cKO mice in the stab-wound injury model (A) Genetic strategy targeting exon 4 of the Chd8 gene. The exon was ﬂanked with loxP sites to excise the loci, resulting in a frameshift mutation that disrupts the production of the CHD8 protein after Cre excision.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies CHD8 Rabbit polyclonal 1 to 500 Abcam ab84527 CHD8 Rabbit polyclonal 1 to 1000 Novus Biologicals NB 100-60418 GFAP Guinea pig polyclonal antiserum 1 to 1000 Synaptic Systems 173 004 Iba1 Chicken polyclonal 1 to 1000 Synaptic Systems 234 006 Iba1 Rabbit Monoclonal 1 to 1000 Abcam ab178846 BrdU Mouse Monoclonal 1 to 200 Sigma-Aldrich MAB4072 Ki67 Rabbit Monoclonal 1 to 500 Invitrogen MA5-14520 tdTomato Rat Monoclonal 1 to 500 Kerafast EST203 LCAT Rabbit polyclonal 1 to 200 for IHC, 1 to 500 for WB Invitrogen PA5-22965 DR3/TNFRSF25 Mouse Monoclonal 1 to 200 for IHC,1 to 500 for WB R&D Systems MAB943 SOX9 Goat antiserum 1 to 200 R&D Systems AF3075 HA tag Rat IgG1 1 to 200 Roche 11867431001 goat-anti-mouse IRDye680 Li-COR Biosciences 926–68070 donkey-anti-rabbit IRDye 800CW Li-COR Biosciences 926–32213 Bacterial and virus strains pAAV2/5 Addgene Addgene # 104964 pAdDeltaF6 helper Addgene Addgene # 112867 pAAV-U6-scramble-GfaABC1D-SaCas9-HA vector This paper N/A pAAV-U6-Rosa26-GfaABC1D-SaCas9-HA vector This paper N/A pAAV-U6-Chd8-GfaABC1D-SaCas9-HA vector This paper N/A Chemicals, peptides, and recombinant proteins Tamoxifen Sigma T5648 OptiPrep Density Gradient Sigma D1556 DAPI Sigma D9542 Antifade mounting medium Vector Laboratories VECTH1000 BrdU Sigma B5002 Lipopolysaccharide Sigma L2880 Cas9 protein NEB M0646T Critical commercial assays RNA synthesis kit NEB E2040S LA-PCR Takara RR002A RNeasy mini kit QIAGEN 74104 iScript cDNA Synthesis Kit Bio-Rad 1725035 iQ SYBR Green Super mix kit Bio-Rad 1708882 TruSeq ribodepleted mRNA stranded for library preparation Illumina 20020594 ATAC-Seq Kit Active Motif 53150 Adult Brain Dissociation Kit, mouse and rat Miltenyi Biotec 130-107-677 Anti-CD11b Magnetic Microbeads kit for microglia Miltenyi Biotec 130–093-634 Anti-ACSA-2 MicroBead Kit for astrocyte Miltenyi Biotec 130-097-678 Deposited data Tissue Bulk RNA-seq data NCBI - Gene Expression Omnibus GEO accession number: GSE271650 Microglia Bulk RNA-seq data NCBI - Gene Expression Omnibus GEO accession number: GSE271651 Astrocyte ATAC-seq data NCBI - Gene Expression Omnibus GEO accession number: GSE271652 (Continued on next page) Cell Reports 43, 114637, August 27, 2024 21

Techniques: Mutagenesis

Figure 2. Reduced reactive gliosis in astrocyte Chd8-cKO mice in the stab-wound injury model (A) Strategy for tamoxifen-induced, astrocyte-speciﬁc Chd8 cKO utilizing the Aldh1l1-CreERT2 line. Mice were crossed with the Ai14-tdTomato reporter line to visualize recombined cells.

Journal: Cell reports

Article Title: Autism-associated CHD8 controls reactive gliosis and neuroinflammation via remodeling chromatin in astrocytes.

doi: 10.1016/j.celrep.2024.114637

Figure Lengend Snippet: Figure 2. Reduced reactive gliosis in astrocyte Chd8-cKO mice in the stab-wound injury model (A) Strategy for tamoxifen-induced, astrocyte-speciﬁc Chd8 cKO utilizing the Aldh1l1-CreERT2 line. Mice were crossed with the Ai14-tdTomato reporter line to visualize recombined cells.

Techniques:

Figure 3. Reduced proliferation and impaired morphological changes of reactive astrocytes in astrocyte Chd8-cKO mice (A and B) Representative images show reduced staining for proliferation markers (BrdU and Ki67) after stab-wound injury in astrocyte cKO mice (B) as compared to control mice (A). The dashed rectangle indicates the ROI used to quantify BrdU+ and Ki67+ cells in (C)–(F). (C) Decreased BrdU+ nuclei in the astrocyte cKO mice as shown by high-magniﬁcation images and quantiﬁcation. (D) Reduced Ki67+ nuclei in the astrocyte cKO mice. (E) Decreased proliferation of tdTomato+ astrocytes from astrocyte cKO mice as shown by reduced tdTomato+/BrdU+ colocalized cells. (F) Decreased proliferation of tdTomato+ astrocytes from astrocyte cKO mice as shown by reduced tdTomato+/BrdU+ colocalized cells. (G and H) Representative skeletonized images of two astrocytes from control (G) and astrocyte cKO mice (H).

Journal: Cell reports

Article Title: Autism-associated CHD8 controls reactive gliosis and neuroinflammation via remodeling chromatin in astrocytes.

doi: 10.1016/j.celrep.2024.114637

Figure Lengend Snippet: Figure 3. Reduced proliferation and impaired morphological changes of reactive astrocytes in astrocyte Chd8-cKO mice (A and B) Representative images show reduced staining for proliferation markers (BrdU and Ki67) after stab-wound injury in astrocyte cKO mice (B) as compared to control mice (A). The dashed rectangle indicates the ROI used to quantify BrdU+ and Ki67+ cells in (C)–(F). (C) Decreased BrdU+ nuclei in the astrocyte cKO mice as shown by high-magniﬁcation images and quantiﬁcation. (D) Reduced Ki67+ nuclei in the astrocyte cKO mice. (E) Decreased proliferation of tdTomato+ astrocytes from astrocyte cKO mice as shown by reduced tdTomato+/BrdU+ colocalized cells. (F) Decreased proliferation of tdTomato+ astrocytes from astrocyte cKO mice as shown by reduced tdTomato+/BrdU+ colocalized cells. (G and H) Representative skeletonized images of two astrocytes from control (G) and astrocyte cKO mice (H).

Techniques: Staining, Control

Figure 6. CHD8 mediates chromatin accessibility changes during LPS-induced neuroinﬂammation (A) Flowchart of the experimental pipeline used to obtain astrocyte-enriched mouse brain samples for the subsequent ATAC-seq processing. (B) Genomic annotation enrichment for altered (increased and decreased) chromatin accessibility between saline-treated control mice and LPS-treated control mice. (C) Genomic annotation enrichment for altered (increased and decreased) chromatin accessibility between LPS-treated astrocyte cKO mice and LPS-treated control mice. (D) Heatmap display of ATAC-seq deviations in chromatin accessibility across the three conditions. (E) Heatmap representation of changes in ATAC-seq peaks near DEGs identiﬁed in our bulk RNA-seq experiments in Figure 5A. (F and G) Representative genome tracks showing loss of accessibility proximal to transcription start sites of the Basp1 gene identiﬁed in a previous study (F) and of the Lcat gene identiﬁed in our RNA-seq (G), in the astrocyte cKO samples.

Journal: Cell reports

Article Title: Autism-associated CHD8 controls reactive gliosis and neuroinflammation via remodeling chromatin in astrocytes.

doi: 10.1016/j.celrep.2024.114637

Figure Lengend Snippet: Figure 6. CHD8 mediates chromatin accessibility changes during LPS-induced neuroinﬂammation (A) Flowchart of the experimental pipeline used to obtain astrocyte-enriched mouse brain samples for the subsequent ATAC-seq processing. (B) Genomic annotation enrichment for altered (increased and decreased) chromatin accessibility between saline-treated control mice and LPS-treated control mice. (C) Genomic annotation enrichment for altered (increased and decreased) chromatin accessibility between LPS-treated astrocyte cKO mice and LPS-treated control mice. (D) Heatmap display of ATAC-seq deviations in chromatin accessibility across the three conditions. (E) Heatmap representation of changes in ATAC-seq peaks near DEGs identiﬁed in our bulk RNA-seq experiments in Figure 5A. (F and G) Representative genome tracks showing loss of accessibility proximal to transcription start sites of the Basp1 gene identiﬁed in a previous study (F) and of the Lcat gene identiﬁed in our RNA-seq (G), in the astrocyte cKO samples.

Techniques: Saline, Control, RNA Sequencing

Figure 7. CRISPR-SaCas9-mediated Chd8 editing through AAV in astrocytes mitigates reactive gliosis in the stab-wound injury model (A) Schematic diagram illustrating the elements required for the designed AAV for astrocyte-speciﬁc Chd8 editing in vivo via CRISPR-SaCas9. (B) Diagram for the simultaneous AAV injection and stab-wound injury with analysis of reactive gliosis performed at 7 days post-injection. (C) Representative images near the needle track from control mice (Scramble-AAV injected). CHD8 is detectable in astrocytes (SOX9+) expressing SaCas9 (HA tag+). (D) Representative images near the needle track from mice injected with the Chd8-cKO AAVs. CHD8 is undetectable in the majority of HA+ and SOX9+ astrocytes (white arrows), while fewer HA+ and SOX9+ astrocytes still show CHD8 expression (yellow arrows). (E and F) GFAP and Iba1 staining of astrocytes and microglia, respectively, after stab-wound injury and AAV injection in the Scramble-AAV (E) and Chd8-cKO-AAV groups (F). (G) Decreased area occupied by GFAP+ astrocytes in the Chd8-cKO-AAV mice. (H) Quantiﬁcation of the area occupied by Iba1+ microglia between control and Chd8-cKO-AAV mice. In (C) and (D), scale bars indicate 20 mm; in (E) and (F), scale bars indicate 500 mm. The dashed rectangle indicates the ROIs that were quantiﬁed in (G) and (H). The dashed lines in merged images indicate the needle track of the injury. In (G) and (H), statistical comparisons were performed with two-way ANOVA; **p < 0.01. Data points indicate n = 3 mice per group. Data are normalized to the means of the ipsilateral site in the control group.

Journal: Cell reports

Article Title: Autism-associated CHD8 controls reactive gliosis and neuroinflammation via remodeling chromatin in astrocytes.

doi: 10.1016/j.celrep.2024.114637

Figure Lengend Snippet: Figure 7. CRISPR-SaCas9-mediated Chd8 editing through AAV in astrocytes mitigates reactive gliosis in the stab-wound injury model (A) Schematic diagram illustrating the elements required for the designed AAV for astrocyte-speciﬁc Chd8 editing in vivo via CRISPR-SaCas9. (B) Diagram for the simultaneous AAV injection and stab-wound injury with analysis of reactive gliosis performed at 7 days post-injection. (C) Representative images near the needle track from control mice (Scramble-AAV injected). CHD8 is detectable in astrocytes (SOX9+) expressing SaCas9 (HA tag+). (D) Representative images near the needle track from mice injected with the Chd8-cKO AAVs. CHD8 is undetectable in the majority of HA+ and SOX9+ astrocytes (white arrows), while fewer HA+ and SOX9+ astrocytes still show CHD8 expression (yellow arrows). (E and F) GFAP and Iba1 staining of astrocytes and microglia, respectively, after stab-wound injury and AAV injection in the Scramble-AAV (E) and Chd8-cKO-AAV groups (F). (G) Decreased area occupied by GFAP+ astrocytes in the Chd8-cKO-AAV mice. (H) Quantiﬁcation of the area occupied by Iba1+ microglia between control and Chd8-cKO-AAV mice. In (C) and (D), scale bars indicate 20 mm; in (E) and (F), scale bars indicate 500 mm. The dashed rectangle indicates the ROIs that were quantiﬁed in (G) and (H). The dashed lines in merged images indicate the needle track of the injury. In (G) and (H), statistical comparisons were performed with two-way ANOVA; **p < 0.01. Data points indicate n = 3 mice per group. Data are normalized to the means of the ipsilateral site in the control group.

Techniques: CRISPR, In Vivo, Injection, Control, Expressing, Staining

TNKS promoted lung adenocarcinoma cell proliferation in vitro (A and C) CCK‐8 and colony formation assays revealed that depletion of TNKS inhibited the growth and proliferation of A549 and H1975 cells. (B and D) TNKS overexpression promoted the growth and proliferation of A549 and H1975 cells. (E) PARylation assays revealed enhanced activity of TNKS‐overexpressing cells and decreased activity of TNKS‐knockdown cells in A549 and H1975 cells. (F ) Western blot assays were adopted to determine the levels of PKA, Duplin, Stbm, β‐catenin, C‐myc, Bcl‐2 and caspase‐3 between the four groups.* p < 0.05, ** p < 0.01.

Journal: Journal of Cellular and Molecular Medicine

Article Title: Study of the mechanism by which Xiaoyan decoction combined with E7449 regulates tumorigenesis in lung adenocarcinoma

doi: 10.1111/jcmm.18467

Figure Lengend Snippet: TNKS promoted lung adenocarcinoma cell proliferation in vitro (A and C) CCK‐8 and colony formation assays revealed that depletion of TNKS inhibited the growth and proliferation of A549 and H1975 cells. (B and D) TNKS overexpression promoted the growth and proliferation of A549 and H1975 cells. (E) PARylation assays revealed enhanced activity of TNKS‐overexpressing cells and decreased activity of TNKS‐knockdown cells in A549 and H1975 cells. (F ) Western blot assays were adopted to determine the levels of PKA, Duplin, Stbm, β‐catenin, C‐myc, Bcl‐2 and caspase‐3 between the four groups.* p < 0.05, ** p < 0.01.

Article Snippet: The primary antibodies used were mouse anti‐TNKS (cat no. ab13587; Abcam, Cambridge, UK), mouse anti‐β‐catenin (cat no. AF6266; Affinity Biosciences), mouse anti‐Myc proto‐oncogene protein (c‐Myc) (cat no. sc‐40; Santa Cruz Biotechnology, Inc. Dallas, TX, USA), rabbit anti‐bcl‐2 (cat no. AF6139; Affinity Biosciences), rabbit anti‐caspase‐3 (cat no. AF6311; Affinity Biosciences), TNKS polyclonal antibody (cat no.18030‐1‐AP, proteintech), rabbit polyclonal anti‐PKA alpha/beta/gamma CAT (cat no. AF7746; Affinity Biosciences), rabbit polyclonal antibody to Duplin (CHD8) (cat no. DF9383; Affinity Biosciences), rabbit polyclonal anti‐stbm (VANGL1) (cat no. DF4607; Affinity Biosciences), rabbit polyclonal anti‐IDUNA (RNF146) (cat no. Bs‐11669R; Bioss), AXIN1 Polyclonal antibody (cat no.16541‐1‐AP, proteintech) and mouse anti‐β‐actin (cat no. MAB8929; OriGene Technologies, Inc.).

Techniques: In Vitro, CCK-8 Assay, Over Expression, Activity Assay, Knockdown, Western Blot

Effects of drug treatment on the protein expression profiles of A549 cells. (A) Venn diagrams revealed a total of 5123 proteins and the differentially expressed proteins in the control group, E7449 group, Xiaoyan decoction group, and Xiaoyan plus E7449 group were 7, 15, 15 and 12, respectively. Furthermore, 4584 differentially expressed proteins coexisted in all groups (A). (B) There were 384, 328 and 280 proteins upregulated in the Xiaoyan decoction, E7449 and combined groups (indicated as X, E and EX in the figure) compared with the control group, respectively. In contrast, 833, 273 and 375 proteins were downregulated in the Xiaoyan decoction, E7449 and combined groups, respectively. (C) GO analysis of the related genes showed that the common differentially expressed genes were associated with the top 20 cellular components, molecular functions, and biological processes. (D) A heatmap was generated for the principal component analysis of all the proteins. (E) A heatmap was generated for the DEPs involved in the WNT signalling pathway. (F, G and H) KEGG analysis showed downregulated proteins in the Wnt pathway in each administration group compared to those in the control group. (I) PPI analysis revealed that TNKS, PKA, Duplin and stbm closely interact with other proteins and corresponded to critical nodes in the network.

Journal: Journal of Cellular and Molecular Medicine

Article Title: Study of the mechanism by which Xiaoyan decoction combined with E7449 regulates tumorigenesis in lung adenocarcinoma

doi: 10.1111/jcmm.18467

Figure Lengend Snippet: Effects of drug treatment on the protein expression profiles of A549 cells. (A) Venn diagrams revealed a total of 5123 proteins and the differentially expressed proteins in the control group, E7449 group, Xiaoyan decoction group, and Xiaoyan plus E7449 group were 7, 15, 15 and 12, respectively. Furthermore, 4584 differentially expressed proteins coexisted in all groups (A). (B) There were 384, 328 and 280 proteins upregulated in the Xiaoyan decoction, E7449 and combined groups (indicated as X, E and EX in the figure) compared with the control group, respectively. In contrast, 833, 273 and 375 proteins were downregulated in the Xiaoyan decoction, E7449 and combined groups, respectively. (C) GO analysis of the related genes showed that the common differentially expressed genes were associated with the top 20 cellular components, molecular functions, and biological processes. (D) A heatmap was generated for the principal component analysis of all the proteins. (E) A heatmap was generated for the DEPs involved in the WNT signalling pathway. (F, G and H) KEGG analysis showed downregulated proteins in the Wnt pathway in each administration group compared to those in the control group. (I) PPI analysis revealed that TNKS, PKA, Duplin and stbm closely interact with other proteins and corresponded to critical nodes in the network.

Techniques: Expressing, Control, Generated

Western blot assays were adopted to compare the levels of PKA, Duplin, Stbm, β‐catenin, C‐myc, Bcl‐2 and caspase‐3 among the four groups.

Journal: Journal of Cellular and Molecular Medicine

Article Title: Study of the mechanism by which Xiaoyan decoction combined with E7449 regulates tumorigenesis in lung adenocarcinoma

doi: 10.1111/jcmm.18467

Figure Lengend Snippet: Western blot assays were adopted to compare the levels of PKA, Duplin, Stbm, β‐catenin, C‐myc, Bcl‐2 and caspase‐3 among the four groups.

Techniques: Western Blot

E7449 or Xiaoyan decoction inhibited lung tumour growth in vivo by regulating the Wnt signalling pathway. (A) Subcutaneous tumour formation experiments in nude mice treated with NaCl ( n = 6), E7449 ( n = 5), Xiaoyan decoction ( n = 4) and E7449 joint Xiaoyan decoction ( n = 4). Representative images of xenograft tumours are shown in the right panel. (B) Tumour volume in the xenograft mice from the treatment group and the control group. (C) Haematoxylin and eosin staining showed that compared to those in the control group, the malignant and aggressive abilities of the E7449 or Xiaoyan decoction group were markedly attenuated. (D) PKA, Duplin, Stbm, β‐catenin, C‐myc, and Bcl‐2 expression was downregulated and caspase‐3 expression was upregulated compared to that in the control group. (E) Electron microscopy image showed ultrastructural alterations in each group. “N” represents the nucleus of the tumour cells in the control group, which were large and irregular. Yellow “*” indicated that the mitochondria were mildly enlarged. In E7449 group, the black arrow shows the cellular autophagy and the white arrow shows that endoplasmic reticulum swelling was observed in the cytoplasm. In Xiaoyan decoction group, the yellow arrow indicated that the chromatin had a tendency of edge aggregation and the black arrow showed mitochondrial autophagy. Furthermore, the yellow arrow indicated that the chromatin was condensed and that edge aggregation occurred; autophagy (black arrow) and mitochondrial swelling (white arrow) were found in the cytoplasm of the group treated with E7449 combined with Xiaoyan decoction group.

Journal: Journal of Cellular and Molecular Medicine

Article Title: Study of the mechanism by which Xiaoyan decoction combined with E7449 regulates tumorigenesis in lung adenocarcinoma

doi: 10.1111/jcmm.18467

Figure Lengend Snippet: E7449 or Xiaoyan decoction inhibited lung tumour growth in vivo by regulating the Wnt signalling pathway. (A) Subcutaneous tumour formation experiments in nude mice treated with NaCl ( n = 6), E7449 ( n = 5), Xiaoyan decoction ( n = 4) and E7449 joint Xiaoyan decoction ( n = 4). Representative images of xenograft tumours are shown in the right panel. (B) Tumour volume in the xenograft mice from the treatment group and the control group. (C) Haematoxylin and eosin staining showed that compared to those in the control group, the malignant and aggressive abilities of the E7449 or Xiaoyan decoction group were markedly attenuated. (D) PKA, Duplin, Stbm, β‐catenin, C‐myc, and Bcl‐2 expression was downregulated and caspase‐3 expression was upregulated compared to that in the control group. (E) Electron microscopy image showed ultrastructural alterations in each group. “N” represents the nucleus of the tumour cells in the control group, which were large and irregular. Yellow “*” indicated that the mitochondria were mildly enlarged. In E7449 group, the black arrow shows the cellular autophagy and the white arrow shows that endoplasmic reticulum swelling was observed in the cytoplasm. In Xiaoyan decoction group, the yellow arrow indicated that the chromatin had a tendency of edge aggregation and the black arrow showed mitochondrial autophagy. Furthermore, the yellow arrow indicated that the chromatin was condensed and that edge aggregation occurred; autophagy (black arrow) and mitochondrial swelling (white arrow) were found in the cytoplasm of the group treated with E7449 combined with Xiaoyan decoction group.

Techniques: In Vivo, Control, Staining, Expressing, Electron Microscopy

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