Journal: Stem Cell Research & Therapy

Article Title: NCAPH-YAP1 interaction promotes breast cancer stemness and tumor progression

doi: 10.1186/s13287-025-04648-0

Figure Lengend Snippet: Transcriptome sequencing analysis of BC cell lines after NCAPH overexpression. A Heatmap of DEGs following NCAPH overexpression. B GSEA analysis showing enrichment of the Hippo signaling pathway. C KEGG pathway analysis of DEGs. D KMplot database analysis of the impact of YAP1 expression levels on OS in BC patients

Article Snippet: The primary antibodies used were against NCAPH (1:5000, Cat# 67,655–1-Ig), CD133 (1:5000, Cat#18,470–1-AP), CD44 (1:5000, Cat#60,224–1-Ig), YAP1 (1:5000, Cat# 13,584–1-AP), phospho-YAP1 (Ser127, 1:5000, Cat#29,018–1-AP), OCT4 (1:5000, Cat#60,242–1-Ig), SOX2 (1:2000, Cat#66,411–1-Ig), Nanog (1:2000, Cat#67,255–1-Ig), Cyclin D1 (1:5000, Cat#26,939–1-AP), CDK1 (1:5000, Cat#67,575–1-Ig), CDK2 (1:5000, Cat#10,122–1-AP), CDK4 (1:5000, Cat#66,950–1-Ig), β-actin (1:5000, Cat#66,009–1-Ig), Flag (1:5000, Cat#66,008–4-Ig), α-Tubulin (1:5000, Cat#66,031–1-Ig), and GAPDH (1:5000, Cat#10,494–1-AP), all sourced from Proteintech (Rosemont, IL, USA).

Techniques: Sequencing, Over Expression, Expressing

Journal: Stem Cell Research & Therapy

Article Title: NCAPH-YAP1 interaction promotes breast cancer stemness and tumor progression

doi: 10.1186/s13287-025-04648-0

Figure Lengend Snippet: NCAPH interacts with YAP1 and promotes YAP1 expression and nuclear translocation. A WB analysis examining the effects of NCAPH overexpression or knockdown on Hippo-YAP1 signaling (Full-length gels are presented in Supplementary Fig. 6). B WB analysis assessing the effect of NCAPH overexpression on YAP1 nuclear translocation (Full-length gels are presented in Supplementary Fig. 7). C IF showing co-localization of NCAPH and YAP1 proteins within cells. D Co-IP assay demonstrating the interaction between NCAPH and YAP1 (Full-length gels are presented in Supplementary Fig. 7)

Article Snippet: The primary antibodies used were against NCAPH (1:5000, Cat# 67,655–1-Ig), CD133 (1:5000, Cat#18,470–1-AP), CD44 (1:5000, Cat#60,224–1-Ig), YAP1 (1:5000, Cat# 13,584–1-AP), phospho-YAP1 (Ser127, 1:5000, Cat#29,018–1-AP), OCT4 (1:5000, Cat#60,242–1-Ig), SOX2 (1:2000, Cat#66,411–1-Ig), Nanog (1:2000, Cat#67,255–1-Ig), Cyclin D1 (1:5000, Cat#26,939–1-AP), CDK1 (1:5000, Cat#67,575–1-Ig), CDK2 (1:5000, Cat#10,122–1-AP), CDK4 (1:5000, Cat#66,950–1-Ig), β-actin (1:5000, Cat#66,009–1-Ig), Flag (1:5000, Cat#66,008–4-Ig), α-Tubulin (1:5000, Cat#66,031–1-Ig), and GAPDH (1:5000, Cat#10,494–1-AP), all sourced from Proteintech (Rosemont, IL, USA).

Techniques: Expressing, Translocation Assay, Over Expression, Knockdown, Co-Immunoprecipitation Assay

Journal: Stem Cell Research & Therapy

Article Title: NCAPH-YAP1 interaction promotes breast cancer stemness and tumor progression

doi: 10.1186/s13287-025-04648-0

Figure Lengend Snippet: The YAP1 inhibitor Verteporfin reverses NCAPH overexpression-induced enhancement of CSC properties, proliferation, migration, and invasion in BC cells. A Verteporfin inhibits YAP1 expression and also reverses the increase in tumorsphere formation induced by NCAPH overexpression (Full-length gels are presented in Supplementary Fig. 8). B WB experiments show that Verteporfin reverses CSC marker protein expression induced by NCAPH overexpression (Full-length gels are presented in Supplementary Fig. 8). C The CCK-8 assay shows that Verteporfin reverses the enhanced proliferative capacity in BC cell lines induced by NCAPH overexpression. D ​The wound healing assay indicates that Verteporfin reverses the increase in migratory capacity of BC cell lines induced by NCAPH overexpression. E The Transwell assay reveals that Verteporfin markedly reverses the enhancement of migratory and invasive capabilities in BC cell lines induced by NCAPH overexpression. Data are presented as mean ± SD (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001)

Article Snippet: The primary antibodies used were against NCAPH (1:5000, Cat# 67,655–1-Ig), CD133 (1:5000, Cat#18,470–1-AP), CD44 (1:5000, Cat#60,224–1-Ig), YAP1 (1:5000, Cat# 13,584–1-AP), phospho-YAP1 (Ser127, 1:5000, Cat#29,018–1-AP), OCT4 (1:5000, Cat#60,242–1-Ig), SOX2 (1:2000, Cat#66,411–1-Ig), Nanog (1:2000, Cat#67,255–1-Ig), Cyclin D1 (1:5000, Cat#26,939–1-AP), CDK1 (1:5000, Cat#67,575–1-Ig), CDK2 (1:5000, Cat#10,122–1-AP), CDK4 (1:5000, Cat#66,950–1-Ig), β-actin (1:5000, Cat#66,009–1-Ig), Flag (1:5000, Cat#66,008–4-Ig), α-Tubulin (1:5000, Cat#66,031–1-Ig), and GAPDH (1:5000, Cat#10,494–1-AP), all sourced from Proteintech (Rosemont, IL, USA).

Techniques: Over Expression, Migration, Expressing, Marker, CCK-8 Assay, Wound Healing Assay, Transwell Assay

Journal: Military Medical Research

Article Title: Endophilin A2 controls touch and mechanical allodynia via kinesin-mediated Piezo2 trafficking.

doi: 10.1186/s40779-024-00520-z

Figure Lengend Snippet: Fig. 9 Hypothetical model illustrating that endophilin A2 (EndoA2) interacts with Piezo2 and KIF5B to form a complex to regulate the membrane trafficking of Piezo2 in sensory neurons and thus contributes to mechanical hypersensitivity

Article Snippet: DRG neurons from L4–L6 of mice were cultured for 2 d. Following fixation, DRG coverslips were treated with primary antibodies against Piezo2 (rabbit, 1:200, Novus, USA, NBP1-78,624) and NF200 (mouse, 1:200, Sigma, USA, N0142) and the appropriate secondary antibodies.

Techniques: Membrane

Journal: Cancers

Article Title: Proteome-Wide Analysis Reveals TFEB Targets for Establishment of a Prognostic Signature to Predict Clinical Outcomes of Colorectal Cancer.

doi: 10.3390/cancers15030744

Figure Lengend Snippet: Figure 1. Mutations in S138 and S142 caused almost complete nuclear retention of TFEB. (A) Schematic of the human TFEB protein domains. S138 and S142 are localized in the proxim- ity of a NES. (B) S138 and S142 of TFEB are evolutionarily conserved in the indicated species. Alignment of the sequences near TFEB S138 and S142 is shown. (C) Western blots of endogenous and exogenous TFEB in CRC cells transfected with wild-type TFEB-GFP or TFEB-GFP mutant (S142A/S138A). Uncropped immunoblots are provided in the Figure S1. (D,E) Localization of TFEBWT and TFEBS142A/S138A in the cytosol and in the nucleus was detected by confocal microscopy. Representative images (D) and statistical results (E) are shown. Torin1 (250 nm), a positive control. Cell outlines in white dotted lines. DAPI was used to label the nucleus. Scale bars, 5 µm. Mean ± SD, n = 10 cells per condition, unpaired t-test. (F) Cell viability of indicated cells were analyzed using a CCK8 assay. Mean ± SEM, n = 3, unpaired t-test. ** p < 0.01, *** p < 0.001.

Article Snippet: The primary antibodies (against TFEB, GFP, LAMP2, NDRG1, SMPD1, and actin) and the HRP-conjugated secondary antibodies (goat anti-rabbit antibody) were supplied by Proteintech (Wuhan, China).

Techniques: Western Blot, Transfection, Mutagenesis, Confocal Microscopy, Positive Control, CCK-8 Assay

Journal: PLoS Genetics

Article Title: Histone Acetyl Transferase 1 Is Essential for Mammalian Development, Genome Stability, and the Processing of Newly Synthesized Histones H3 and H4

doi: 10.1371/journal.pgen.1003518

Figure Lengend Snippet: A) Schematic diagram of the wild type mouse Hat1 locus (top), the Hat1 locus following integration of loxP sequences flanking intron three (middle) and the Hat1 locus following Cre mediated deletion of exon 3. Exons are represented by purple rectangles. Locations of probes and PCR primers are indicated. (B) Genomic DNA isolated from a parental (C57/bl6) mouse and mice generated from a cross between a chimeric mouse and a wild type mouse was digested with EcoRV and analyzed by Southern blot using the indicated probe. Mice 1, 4 and 6 are Hat1 flox/WT . C) A Hat1 flox/WT mouse was crossed with a mouse that ubiquitously expresses the cre recombinase. Genomic DNA was isolated from mice generated by this cross, digested with EcoRV and analyzed by Southern blot with the indicated probe. Mice 3, 5 and 9 are WT/KO/Cre. D) Table lists the number of expected, obtained and viable pups of the indicated genomes derived from matings of Hat1 +/− mice. E) PCR genotyping of a representative litter from a Hat1 +/− X Hat1 +/− mating. Primers P1 and P2 (see above) were used for amplification. Arrows indicate specific PCR products. F) Representative neonatal pups from Hat1 +/− X Hat1 +/− matings with the indicated genotype. F) Body weight of pups was measured immediately following birth. Data are derived from 10 pups of each genotype.

Article Snippet: Slides were also stained with primary antibodies against Hat1 (Abcam, 1/50 dilution), α-Ki67 (Novocastra, NCL-KI67-P) and anti-Cleaved Caspase 3 (Cell Signaling, #9661)Images were captured with a Zeiss AxioImager Z1 microscope.

Techniques: Isolation, Generated, Southern Blot, Derivative Assay, Amplification

Journal: PLoS Genetics

Article Title: Histone Acetyl Transferase 1 Is Essential for Mammalian Development, Genome Stability, and the Processing of Newly Synthesized Histones H3 and H4

doi: 10.1371/journal.pgen.1003518

Figure Lengend Snippet: A) Histologic appearance of lungs from newborn pups obtained from a Hat1 +/−+ and Hat1 −/− mice. Staining was with hematoxylin-eosin; magnification 20×, (inlets ×40). The lungs of Hat1 −/− show less aeration, due to thickened mesenchyme resulting in death due to respiratory failure. B) Hat1 is highly expressed in lungs of Hat1 +/+ but not in Hat1 −/− mice; magnification 20×, (inlets ×40) C) Cleaved Caspase3 stained by IHC showed no difference between lungs of Hat1 ++ and Hat1 −/− mice; magnification 20×, (inlets ×40) D) Ki67 stained by IHC shows significantly higher proliferation rates in lungs of Hat1 −/− mice compared to controls; magnification 20×, (inlets ×40). Quantification was done by HistoQuest software.

Article Snippet: Slides were also stained with primary antibodies against Hat1 (Abcam, 1/50 dilution), α-Ki67 (Novocastra, NCL-KI67-P) and anti-Cleaved Caspase 3 (Cell Signaling, #9661)Images were captured with a Zeiss AxioImager Z1 microscope.

Techniques: Staining, Software

Journal: PLoS Genetics

Article Title: Histone Acetyl Transferase 1 Is Essential for Mammalian Development, Genome Stability, and the Processing of Newly Synthesized Histones H3 and H4

doi: 10.1371/journal.pgen.1003518

Figure Lengend Snippet: A) Micro CT scans of the heads of neonates with the indicated genotype. Top row shows a dorsal view and bottom row shows a ventral view. Arrows indicate defects in the nasal cavity (top) and jaw structures (bottom). B) Micro CT scans of Hat1 +/+ and Hat1 −/− neonates (as indicated) showing a dorsal view of the entire animal. C) Acian blue and Alizarin red stained Hat1 +/+ and Hat1 −/− neonates. D) Hat1 +/+ and Hat1 −/− embryos (12.5 dpc) were stained with α-Hat1 antibodies. E) Cross section of the head and neck of 11.5 dpc Hat1 +/+ and (WT) and Hat1 −/− embryos stained with α-Hat1 antibody. 2.5× magnification.

Article Snippet: Slides were also stained with primary antibodies against Hat1 (Abcam, 1/50 dilution), α-Ki67 (Novocastra, NCL-KI67-P) and anti-Cleaved Caspase 3 (Cell Signaling, #9661)Images were captured with a Zeiss AxioImager Z1 microscope.

Techniques: Micro-CT, Staining

Journal: PLoS Genetics

Article Title: Histone Acetyl Transferase 1 Is Essential for Mammalian Development, Genome Stability, and the Processing of Newly Synthesized Histones H3 and H4

doi: 10.1371/journal.pgen.1003518

Figure Lengend Snippet: A) Hat1 +/+ , Hat1 +/− and Hat1 −/− primary MEFs were genotyped by PCR as described in the legend to . Whole cell extracts from the indicated MEFs were analyzed by Western blots probed with the indicated antibodies. B) Equal numbers of primary MEFs of the indicated genotype were seeded at time zero. Cell numbers were counted at the indicated time points. C) Primary Hat1 +/+ and Hat1 −/− MEFs were stained with propidium iodide and analyzed by FACS. Fraction of cells in each phase of the cell cycle is indicated. D) Immortalized Hat1 +/+ and Hat1 −/− MEFs were grown under the indicated conditions. Plates were photographed after crystal violet staining.

Article Snippet: Slides were also stained with primary antibodies against Hat1 (Abcam, 1/50 dilution), α-Ki67 (Novocastra, NCL-KI67-P) and anti-Cleaved Caspase 3 (Cell Signaling, #9661)Images were captured with a Zeiss AxioImager Z1 microscope.

Techniques: Western Blot, Staining

Journal: PLoS Genetics

Article Title: Histone Acetyl Transferase 1 Is Essential for Mammalian Development, Genome Stability, and the Processing of Newly Synthesized Histones H3 and H4

doi: 10.1371/journal.pgen.1003518

Figure Lengend Snippet: A) Hat1 +/+ and Hat1 −/− MEFs were stained with either DAPI or α-γH2AX antibodies as indicated (left). Visible γH2AX foci were counted in 12 cells of each genotype (right). B) Metaphase spreads from Hat1 +/+ and Hat1 −/− MEFs were analyzed for chromosome number and the presence of breaks and fusions. The percentage of cells containing the indicated chromosomal abnormality is given. The number of spreads analyzed was 56 (Hat1 +/+ ) and 112 (Hat1 −/− ). C) Metaphase spreads were generated from Hat1 +/+ and Hat1 −/− MEFs. Insets show enlarged views of selected abnormal chromosomes. Red arrows indicate examples of chromosome fusions and blue arrows indicate chromosome breaks.

Article Snippet: Slides were also stained with primary antibodies against Hat1 (Abcam, 1/50 dilution), α-Ki67 (Novocastra, NCL-KI67-P) and anti-Cleaved Caspase 3 (Cell Signaling, #9661)Images were captured with a Zeiss AxioImager Z1 microscope.

Techniques: Staining, Generated

Journal: PLoS Genetics

Article Title: Histone Acetyl Transferase 1 Is Essential for Mammalian Development, Genome Stability, and the Processing of Newly Synthesized Histones H3 and H4

doi: 10.1371/journal.pgen.1003518

Figure Lengend Snippet: A and B) Hat1 +/+ and Hat1 −/− MEFs were pulse-labeled with EdU and chased with thymidine as schematically depicted at the top of each panel. Samples were isolated at the indicated time points and proteins associated with nascent DNA were resolved by SDS-PAGE following affinity purification of EdU-labeled DNA (iPond). The indicated amounts of the input fractions (prior to affinity purification) are on the left-hand side of each panel. Samples eluted from the affinity purification resin are on the right-hand side of each panel. In all cases, the respective Hat1 +/+ and Hat1 −/− samples were analyzed on the same gel (separated by a MW standard lane that has been removed). Western blots were probed with the antibodies indicated on the right. Numbers below each lane indicate the normalized intensity of the band as determined using Licor software. Each sample was normalized to the level of unmodified histone H3. The intensity observed in the 30′ pulse sample was arbitrarily set to 1.0. No Clck indicates control samples that were not biotin labeled.

Article Snippet: Slides were also stained with primary antibodies against Hat1 (Abcam, 1/50 dilution), α-Ki67 (Novocastra, NCL-KI67-P) and anti-Cleaved Caspase 3 (Cell Signaling, #9661)Images were captured with a Zeiss AxioImager Z1 microscope.

Techniques: Labeling, Isolation, SDS Page, Affinity Purification, Western Blot, Software

Journal: PLoS Genetics

Article Title: Histone Acetyl Transferase 1 Is Essential for Mammalian Development, Genome Stability, and the Processing of Newly Synthesized Histones H3 and H4

doi: 10.1371/journal.pgen.1003518

Figure Lengend Snippet: Hat1 +/+ and Hat1 −/− MEFs were pulse-labeled with 3 H-lysine for 12 minutes. Histones were then isolated and resolved by Acid-Urea (AU) gel electrophoresis. Total protein was visualized with Coomassie blue staining and radio-labeled proteins visualized by fluorography (as indicated). The mobility of each histone is indicated. The brackets indicate the regions of mobility for the acetylated isoforms of histone H4 and histone H3.

Article Snippet: Slides were also stained with primary antibodies against Hat1 (Abcam, 1/50 dilution), α-Ki67 (Novocastra, NCL-KI67-P) and anti-Cleaved Caspase 3 (Cell Signaling, #9661)Images were captured with a Zeiss AxioImager Z1 microscope.

Techniques: Labeling, Isolation, Nucleic Acid Electrophoresis, Staining

Journal: Open Medicine

Article Title: Luteolin alleviates ulcerative colitis in rats via regulating immune response, oxidative stress, and metabolic profiling

doi: 10.1515/med-2023-0785

Figure Lengend Snippet: Effects of LUT on inflammation and immune response in UC rats. (a) Expression levels of TNF-α, CRP, IL-13, IL-33, and ST-2 were examined by ELISA. (b) Representative image of CD4+ T cell staining in the colon (100× magnification). (c) Representative image of CD8+ T cell staining (100× magnification). NC represents normal control. MOD represents model. MES represent mesalazine. LUT represents luteolin. There were 10, 8, 8, and 9 replicates in NC, MOD, MES, and LUT groups, respectively. ** P < 0.01 vs the NC group. # P < 0.05 and ## P < 0.01 vs the MOD group.

Article Snippet: The sections were blocked with 5% BSA and incubated with primary antibody against CD4 (1:2,000; Abcam, UK) and CD8 (1:2,000; Abcam, UK) overnight at 4°C.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Staining