Journal: Nature chemical biology

Article Title: Membrane editing with proximity labeling reveals regulators of lipid homeostasis

doi: 10.1038/s41589-025-02104-x

Figure Lengend Snippet: a , b , Quantification of miRFP-Nir2 (T59A mutant with lipid transfer activity disabled) co-localization with PA-fed membranes. Black horizontal bars indicate means and vertical error bars indicate standard deviations. Statistical significance was determined using two-sided Student’s t-test ( n = 10–16 cells with exact numbers provided in the Source Data, p = 8.6E-06 and 8.9E-07). c , Confocal images of HEK 293T cells stably expressing V5-tagged Nir2 (PITPNM1), PDZD8, ORP1L (OSBPL1A), full-length SCP2 (SCP-x), or the mature form of SCP2 (mSCP2). Cells were co-stained with α-LAMP1 and α-Calnexin antibodies as lysosome and ER markers. Shown are representative images from two independent experiments. d , e , Zoomed-in images of cells expressing V5-SCP-x ( d ) or V5-mSCP2 ( e ). V5-mSCP2 shows more cytosolic localization. Scale bars, 20 μm. f , Scheme of IMPACT labeling to quantify superPLD activity in live cells. g , h , Flow cytometry results of cells co-expressing superPLD targeted to the plasma membrane (PM; g ) or lysosomes (Lyso; h ) and the indicated lipid-transfer protein. mCherry signal (readout of superPLD expression level) and BODIPY signal (readout of PLD activity) are plotted. Wild-type cells expressing deadPLD were included as a negative control. Shown are representative plots from two independent experiments.

Article Snippet: Antibody and dilutions used for validation experiments ( – ) were the following: anti-LPIN1 polyclonal antibody (Proteintech, 27026–1-AP; 1:1,000), anti-LPIN2 monoclonal antibody (Santa Cruz Biotechnology, sc-514353; 1:100), anti-Nir2 polyclonal antibody (Proteintech, 26983–1-AP; 1:1,000), anti-PDZD8 polyclonal antibody (Proteintech, 25512–1-AP; 1:1,000), anti-SCP-x polyclonal antibody (Proteintech, 14397–1-AP; 1:1,000), anti-SCP-2 polyclonal antibody (Proteintech, 23006–1-AP; 1:1,000), anti-DGKD polyclonal antibody (Abcepta, AP8126b; 1:1,000), anti-DGKH polyclonal antibody (Proteintech, 13873–1-AP; 1:1,000) and anti-β-tubulin monoclonal antibody (Cell Signaling Technology, 86298).

Techniques: Mutagenesis, Activity Assay, Stable Transfection, Expressing, Staining, Labeling, Flow Cytometry, Clinical Proteomics, Membrane, Negative Control

Journal: Nature chemical biology

Article Title: Membrane editing with proximity labeling reveals regulators of lipid homeostasis

doi: 10.1038/s41589-025-02104-x

Figure Lengend Snippet: a , Western blot of LOVPLD-expressing HEK 293T cells treated with DsiRNA to deplete Nir2, SCP-x/SCP2, or PDZD8. DsiRNA #1 (colored red) was used in the subsequent experiments. , Quantification of ( a ), with relative levels compared to the control shown in parentheses. Plots are representative of two independent experiments. c , d , Expression levels of LOVPLD ( c ) and the PA probe GFP-PASS ( d ) in HEK 293T cells depleted with Nir2 (siNir2), SCP2 (siSCP2), or PDZD8 (siPDZD8), co-expressing PA probe and LOVPLD targeted to either plasma membrane (PM) or lysosomes (Lyso). Plots are representative of two independent experiments. e , f , Quantification of colocalization between LOVPLD and GFP-PASS shown in , . Black horizontal bars indicate means and vertical error bars indicate standard deviations. Statistical analysis was performed using one-way ANOVA followed by post-hoc Tukey-HSD test ( n = 14–19 cells with exact numbers provided in the Source Data, p = 0.01 and 0.6 for ( e ) and 0.03 and 0.9 for ( f ) for Control vs. siRNA samples). g , Western blot of p-S6K to measure mTOR activity in LTP-depleted cells expressing LOVPLD, with quantification presented in . LOVPLD was localized to plasma membrane (PM), lysosomes (Lyso), or endoplasmic reticulum (ER), and 30-min incubation with intermittent blue light illumination (470 nm, 500 ms per 5 s) was used to activate PA production by LOVPLD. β-actin was used as a loading control. h , Similar experiment as ( g ) but in LTP-depleted HEK 293T cells without LOVPLD activation. i , Quantification of ( h ). n = 4 (except for siPDZD8, where n = 2) biological replicates. Statistical analysis was performed using one-way ANOVA followed by post-hoc Tukey-HSD test ( p = 0.004, 0.01, and 0.07 for Control vs. siRNA samples). Black horizontal bars indicate means and vertical error bars indicate standard deviations.

Article Snippet: Antibody and dilutions used for validation experiments ( – ) were the following: anti-LPIN1 polyclonal antibody (Proteintech, 27026–1-AP; 1:1,000), anti-LPIN2 monoclonal antibody (Santa Cruz Biotechnology, sc-514353; 1:100), anti-Nir2 polyclonal antibody (Proteintech, 26983–1-AP; 1:1,000), anti-PDZD8 polyclonal antibody (Proteintech, 25512–1-AP; 1:1,000), anti-SCP-x polyclonal antibody (Proteintech, 14397–1-AP; 1:1,000), anti-SCP-2 polyclonal antibody (Proteintech, 23006–1-AP; 1:1,000), anti-DGKD polyclonal antibody (Abcepta, AP8126b; 1:1,000), anti-DGKH polyclonal antibody (Proteintech, 13873–1-AP; 1:1,000) and anti-β-tubulin monoclonal antibody (Cell Signaling Technology, 86298).

Techniques: Western Blot, Expressing, Control, Clinical Proteomics, Membrane, Activity Assay, Incubation, Activation Assay

Journal: The Journal of Biological Chemistry

Article Title: Sterol carrier protein-2 deficiency attenuates diet-induced dyslipidemia and atherosclerosis in mice

doi: 10.1074/jbc.RA118.002290

Figure Lengend Snippet: SCP2 deficiency significantly attenuates diet-induced atherosclerosis. At 10 weeks of age, LDLR−/− and LS mice were fed a high-fat, high-cholesterol–containing Western-type diet (TD88137) for 16 weeks. Aortae were dissected and prepared for en face analyses. The total area as well as the area occupied by the lesions was quantified using Axiovision software and expressed as percent lesion area. A, representative images. B, percent area occupied by the lesions in the aortic arch. C, percent area occupied by the lesions in the entire aorta. *, p < 0.05; individual p values are included in the text.

Article Snippet: Quantitative RT-PCR was used to measure mRNA levels using a TaqMan assay (Hs00920780_m1), and the protein levels were measured by Western blot analyses using primary antibody 23006-1-AP (Proteintech Group), which recognizes both SCP2 and SCPx.

Techniques: Western Blot, Software

Journal: The Journal of Biological Chemistry

Article Title: Sterol carrier protein-2 deficiency attenuates diet-induced dyslipidemia and atherosclerosis in mice

doi: 10.1074/jbc.RA118.002290

Figure Lengend Snippet: SCP2 deficiency reduces plaque size as well as plaque necrosis in the aortic root. Paraffin-embedded aortic root sections (5 μm) were stained with H&E or Masson's trichrome, imaged, and analyzed by Axiovision software. A, representative H&E stained images of the aortic root of the indicated genotypes/sex. Scale bar = 100 μm. B, the total aortic root and area occupied by the lesions was quantified, and data (mean ± S.D., n = 6) are presented as percent lesion area. C, representative trichrome-stained images of the indicated genotypes/sex. Scale bar = 50 μm. D, the total and necrotic areas were quantified for all three aortic valve leaflets, and data (mean ± S.D., n = 12 leaflets) are presented as percent necrotic area. *, p < 0.05.

Article Snippet: Quantitative RT-PCR was used to measure mRNA levels using a TaqMan assay (Hs00920780_m1), and the protein levels were measured by Western blot analyses using primary antibody 23006-1-AP (Proteintech Group), which recognizes both SCP2 and SCPx.

Techniques: Staining, Software

Journal: The Journal of Biological Chemistry

Article Title: Sterol carrier protein-2 deficiency attenuates diet-induced dyslipidemia and atherosclerosis in mice

doi: 10.1074/jbc.RA118.002290

Figure Lengend Snippet: SCP2 deficiency significantly reduces plasma cholesterol and triglyceride levels. At 10 weeks of age, LDLR−/− and LS mice were fed a high-fat, high-cholesterol–containing Western-type diet (TD88137) for 16 weeks. After an overnight fast, mice were euthanized, and fasting plasma was collected. A, levels of total plasma cholesterol for indicated genotype and sexes. B, percent of total plasma cholesterol associated with the non-HDL or HDL fraction. C, percent of total plasma cholesterol associated with the non-HDL or HDL fraction in both genotypes normalized to total plasma cholesterol in LDLR−/− mice of the respective sex. D, linear regression analyses of total lesion area and plasma cholesterol; the observed coefficient of correlation (R) as significance of correlation (p value) is indicated. E, total plasma triglyceride for the indicated genotype and sexes. F, linear regression analyses of total lesion area and plasma triglyceride levels; the observed coefficients of correlation (R) as significance of correlation (p value) is indicated.

Article Snippet: Quantitative RT-PCR was used to measure mRNA levels using a TaqMan assay (Hs00920780_m1), and the protein levels were measured by Western blot analyses using primary antibody 23006-1-AP (Proteintech Group), which recognizes both SCP2 and SCPx.

Techniques: Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Sterol carrier protein-2 deficiency attenuates diet-induced dyslipidemia and atherosclerosis in mice

doi: 10.1074/jbc.RA118.002290

Figure Lengend Snippet: SCP2 deficiency significantly reduces intestinal cholesterol absorption, and SCP2 knockdown in intestinal epithelial cells reduces cholesterol uptake. A, C57BL/6 (WT) or SCP2−/− mice were injected intravenously with the lipoprotein lipase inhibitor tyloxapol (500 mg/kg body weight) and gavaged with [3H]cholesterol in olive oil (2 μCi in 200 μl) after 5 min. Intestinal cholesterol absorption was assessed by monitoring the radiolabel associated with plasma collected at the time of euthanasia. Data are presented as plasma DPM per microliter of plasma for the indicated genotype and sex. B, the entire length of the intestine, from the base of the stomach to the tip of the cecum, was divided into four equal parts (P1 to P4), and total RNA was isolated. The mRNA levels of indicated genes were determined by real-time PCR as described under “Experimental procedures” and normalized to the housekeeping gene β-actin. C, total protein extracts from ileum segments P1 to P4 of WT and SCP2−/− mice as well as the colon (C) were analyzed by Western blotting for expression of SCP2; β-actin was used as the loading control. Human intestinal epithelial cells (HT-29) were transfected with scrambled or SCP2-specific siRNA as described under “Experimental procedures.” Total protein or RNA was extracted and used to determine the levels of SCP2. D, top, a representative Western blot. Bottom, levels of SCP2 mRNA quantified by quantitative PCR and SCP2 protein by densitometric analyses of Western blots. Data are presented as percent scrambled siRNA-transfected controls for the indicated SCP2 siRNA concentrations used. E, HT-29 cells transfected with scrambled siRNA or 53.3 nm SCP2-specific siRNA were incubated with [3H]-cholesterol, and total cellular uptake was monitored as described under “Experimental procedures.” Data (mean ± S.D., n = 6) are presented as DPM normalized to total cellular protein. *, p < 0.05.

Article Snippet: Quantitative RT-PCR was used to measure mRNA levels using a TaqMan assay (Hs00920780_m1), and the protein levels were measured by Western blot analyses using primary antibody 23006-1-AP (Proteintech Group), which recognizes both SCP2 and SCPx.

Techniques: Injection, Isolation, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Transfection, Incubation

Journal: The Journal of Biological Chemistry

Article Title: Sterol carrier protein-2 deficiency attenuates diet-induced dyslipidemia and atherosclerosis in mice

doi: 10.1074/jbc.RA118.002290

Figure Lengend Snippet: SCP2 deficiency significantly reduces lipid secretion from liver and isolated hepatocytes. A, C57BL/6 (WT) or SCP2−/− mice were injected with the lipoprotein lipase inhibitor tyloxapol (500 mg/kg of body weight), and blood samples were drawn at 0 and 3 h. Triglyceride secretion rates for indicated genotypes and sexes are presented. B and C, primary hepatocytes were prepared from C57BL/6 (WT) or SCP2−/− mice. Following incubation with [3H]oleic acid, radiolabel associated with secreted triglycerides (TG, B) or cholesteryl esters (C) was determined as described under “Experimental procedures” and normalized to cellular protein. Data are presented as DPM associated with the triglyceride or cholesteryl ester fraction in the total lipids extracted from the medium per milligram of total protein.

Article Snippet: Quantitative RT-PCR was used to measure mRNA levels using a TaqMan assay (Hs00920780_m1), and the protein levels were measured by Western blot analyses using primary antibody 23006-1-AP (Proteintech Group), which recognizes both SCP2 and SCPx.

Techniques: Isolation, Injection, Incubation

Journal: The Journal of Biological Chemistry

Article Title: Sterol carrier protein-2 deficiency attenuates diet-induced dyslipidemia and atherosclerosis in mice

doi: 10.1074/jbc.RA118.002290

Figure Lengend Snippet: SCP2 deficiency leads to reduced lipid accumulation in the liver without a change in the expression of lipogenic genes. A, liver tissue harvested from WD-fed LDLR−/− or LS mice were paraffin-embedded, and 5-μm sections were stained with H&E. Images were acquired using a Zeiss inverted microscope fitted with a digital camera. Scale bar = 50 μm. B, SCP2 mRNA levels in total liver RNA were determined by real-time PCR as described under “Experimental procedures” and normalized to the housekeeping gene β-actin. Relative expression (mean ± S.D., n = 6) is shown. C, hepatic mRNA levels of the indicated genes were determined by real-time PCR as described under “Experimental procedures” and normalized to the housekeeping gene β-actin. Relative expression (mean ± S.D., n = 6) is shown.

Article Snippet: Quantitative RT-PCR was used to measure mRNA levels using a TaqMan assay (Hs00920780_m1), and the protein levels were measured by Western blot analyses using primary antibody 23006-1-AP (Proteintech Group), which recognizes both SCP2 and SCPx.

Techniques: Expressing, Staining, Inverted Microscopy, Real-time Polymerase Chain Reaction

Journal: The Journal of Biological Chemistry

Article Title: Sterol carrier protein-2 deficiency attenuates diet-induced dyslipidemia and atherosclerosis in mice

doi: 10.1074/jbc.RA118.002290

Figure Lengend Snippet: Effects of SCP2 deficiency on cholesterol accumulation in and cholesterol efflux from macrophages. Thioglycolate-elicited macrophages were isolated from C57BL/6 (WT) and SCP2−/− mice and incubated with AcLDL (25 μg/ml) for 48 h. A and B, following two washes with PBS, cells were either fixed and stained with Oil Red O (A) or used for total lipid extraction and cholesterol mass measurement (B). Total cholesterol mass was normalized to total cellular protein, and data (mean ± S.D., n = 6) are presented as nanomoles per milligram of protein. C, total protein extracts of macrophages were subjected to Western blot analyses to assess SR-A expression; β-actin was used as a loading control. D, for measurement of cholesterol efflux, cells were loaded with AcLDL and labeled with [3H]-cholesterol for 48 h. Following a 24-h equilibration, cholesterol efflux to 10% FBS in the growth medium was monitored over time. Data (mean ± S.D., n = 6) are presented as percent efflux. *, p < 0.05.

Article Snippet: Quantitative RT-PCR was used to measure mRNA levels using a TaqMan assay (Hs00920780_m1), and the protein levels were measured by Western blot analyses using primary antibody 23006-1-AP (Proteintech Group), which recognizes both SCP2 and SCPx.

Techniques: Isolation, Incubation, Staining, Mass Measurement, Western Blot, Expressing, Labeling

Journal: The Journal of Biological Chemistry

Article Title: Sterol carrier protein-2 deficiency attenuates diet-induced dyslipidemia and atherosclerosis in mice

doi: 10.1074/jbc.RA118.002290

Figure Lengend Snippet: Effects of SCP2 deficiency on biliary bile acid and cholesterol secretion. A and B, gall bladder bile was collected at the time of euthanasia, and the total volume was noted. Biliary bile acids (BA), cholesterol, and phospholipids (PL) were estimated as described under “Experimental procedures.” Data are presented as total bile acids (nanomoles) or FC (micrograms) in the bile normalized to total phospholipids (micrograms) in A and B, respectively.

Article Snippet: Quantitative RT-PCR was used to measure mRNA levels using a TaqMan assay (Hs00920780_m1), and the protein levels were measured by Western blot analyses using primary antibody 23006-1-AP (Proteintech Group), which recognizes both SCP2 and SCPx.

Techniques:

Journal: Nature Communications

Article Title: Regulation of NSL by TAF4A is critical for genome stability and quiescence of muscle stem cells

doi: 10.1038/s41467-025-64402-1

Figure Lengend Snippet: a Heat maps of centered TAF4A CUT&RUN peaks. The blue-to-red gradient indicates high-to-low counts in the corresponding region. b Genome-wide distribution of TAF4A peaks. c Bar plot representing TAF4A peaks at different types of promoters. d Detected SP1 and NF-YA binding site under TAF4A CUT&RUN peaks. e Taf4a and Nf-Ya expression in freshly isolated MuSCs (fiMuSCs) and muscle based on reads per exon kilobase per million (RPKM) values (unpaired two-tailed t test: * p = 0.0103, *** p = 0.0005 n = 3). f Co-IP of TAF4A and NF-YA/B in wildtype MuSCs. Blots were probed with antibodies against TAF4A and NF-YA or NF-YB ( n = 2). g Percentage of TAF4A CUT&RUN peaks overlapping either with NF-YA peaks or with SP1 binding motifs. h Venn diagram of overlapping TAF4A CUT&RUN (CnR) peaks, NF-YA CUT&RUN (CnR) peaks, and RNA-seq DEG in Taf4a sKO MuSCs. i TAF4A, NF-YA and H3K4me3 distribution in the proximal promoter region of the Kansl2 gene in WT MuSCs. 8-20-weeks-old males and females were used. Data are presented as mean ± SEM of biological replicates. Source data are provided in the Source Data file.

Article Snippet: In brief, 1 × 10 6 wild type in vitro cultured muscle stem cells (MuSCs) or 4 × 10 5 wild type and Taf4a sKO MuSCs were immobilized on Concanavalin A-coated magnetic beads (Bangs Laboratories), permeated with 0.05% Digitonin (EMD Millipore), and incubated with TAF4A (Santa Cruz #sc-136093), KANSL2 (Proteintech #27261-1-AP), NF-YA (Santa Cruz #sc-17753) or mouse IgG (Millipore #12-371B) or rabbit IgG (Diagenode #C15410206) on a rotator with 1:100 dilution at 4 °C overnight on a rotator.

Techniques: Genome Wide, Binding Assay, Expressing, Isolation, Two Tailed Test, Co-Immunoprecipitation Assay, RNA Sequencing

Journal: Nature Communications

Article Title: Regulation of NSL by TAF4A is critical for genome stability and quiescence of muscle stem cells

doi: 10.1038/s41467-025-64402-1

Figure Lengend Snippet: a Representation of NSL complex functions. b Heat map of expression levels of NSL complex components in Taf4a sKO MuSCs compared to control MuSCs ( n = 3). c , d RT-qPCR analysis of Kansl2 ( c ) and Mcrs1 ( d ) expression in freshly isolated control and Taf4a sKO MuSCs. m36b4 was used as reference gene (unpaired two-tailed t test, ( c ) * p = 0.0161, ( d ) * p = 0.0417, n = 5). e Western blot analysis of H4K16ac in cultured MuSCs from control and Taf4a sKO mice. H3 was used as loading control (unpaired two-tailed t test: * p = 0.0280, n = 3). f Heat maps of CUT&RUN signals of KANSL2 at transcriptional start sites. The blue-to-red gradient indicates high-to-low counts in the corresponding region. g Genome-wide distribution of KANSL2 peaks. h Venn diagram of overlapping KANSL2 CUT&RUN peaks, up and down-regulated genes from RNA-seq data in Taf4a sKO MuSCs. i Heat map showing KANSL2 binding at promoters of downregulated epigenetic modifiers genes in Taf4a sKO MuSCs ( n = 3). j GO term analysis of genes with an overlap between KANSL2 CUT&RUN peaks and downregulated genes in Taf4a sKO MuSCs based on P-values using EnrichR (Fisher’s exact test, two-sided). The number of genes associated with each GO term is indicated in square brackets. 8–20-week-old males and females were used. MuSCs were isolated 4–6 weeks after tamoxifen administration. Data are presented as mean ± SEM of biological replicates. Source data are provided in the Source Data file.

Article Snippet: In brief, 1 × 10 6 wild type in vitro cultured muscle stem cells (MuSCs) or 4 × 10 5 wild type and Taf4a sKO MuSCs were immobilized on Concanavalin A-coated magnetic beads (Bangs Laboratories), permeated with 0.05% Digitonin (EMD Millipore), and incubated with TAF4A (Santa Cruz #sc-136093), KANSL2 (Proteintech #27261-1-AP), NF-YA (Santa Cruz #sc-17753) or mouse IgG (Millipore #12-371B) or rabbit IgG (Diagenode #C15410206) on a rotator with 1:100 dilution at 4 °C overnight on a rotator.

Techniques: Expressing, Control, Quantitative RT-PCR, Isolation, Two Tailed Test, Western Blot, Cell Culture, Genome Wide, RNA Sequencing, Binding Assay

Journal: Nature Communications

Article Title: Regulation of NSL by TAF4A is critical for genome stability and quiescence of muscle stem cells

doi: 10.1038/s41467-025-64402-1

Figure Lengend Snippet: a DAPI staining of Taf4a sKO MuSCs. Quantification of relative micronuclei numbers. Scale bar: 5 µm (**** p < 0.0001, n = 6). b gH2A.X immunofluorescence in cultured control and Taf4a sKO MuSCs. Scale bar: 20 µm. Quantification of relative numbers of γH2A.X+ cells (*** p = 0.0008, n = 4). c Western blot of γH2A.X in control and Taf4a sKO MuSCs. Loading control: H3 ( n = 3). d Western blot of immunoprecipitated Lamin A/C in control and Taf4a sKO MuSCs. Acetylation signals were normalized to total Lamin A/C levels. Quantification of acetylation levels (*** p = 0.0006, n = 3). e Western blot of p-lamin A/C ser392 in cultured control and Taf4a sKO MuSCs. Loading control: GAPDH ( n = 2). f Stiffness measurements of control and Taf4a sKO MuSC nuclei by AFM. Data represent average medians of Young’s Modulus (Pa) normalized to control MuSCs (** p = 0.0017, n = 5). g RT-qPCR analysis of Kansl2 expression in scrambled control and Nf-ya knockdown MuSCs. Reference gene: m36b4 (*** p = 0.0004, *** p = 0.0007, n = 6). h LaminB staining and quantification of micronuclei-positive cells in untreated ( n = 6), scrambled control ( n = 3), Taf4a knockdown ( n = 3), Kansl2 knockdown ( n = 3), Nfya knockdown ( n = 3), and Mcrs1 knockdown ( n = 3) in cultured MuSCs (* p = 0.0214, *** p = 0.0001, **** p < 0.0001). Scale bar: 5 µm. i LaminB staining and quantification of micronuclei-positive cells in control, Kansl2 overexpressing control, Taf4a sKO and Kansl2 overexpressing Taf4a sKO MuSCs (** p = 0.0054 *** p = 0.0004 **** p < 0.0001, n = 4). Scale bar: 5 µm. j Stiffness measurements of control, Taf4a sKO and Kansl2 overexpressing Taf4a sKO MuSC nuclei by AFM. Data representation as in ( f ) (** p = 0.0026, * p = 0.0437 n = 4). k RT-qPCR analysis of Kansl2 expression in scrambled control, Nf-ya knockdown and Nf-ya knockdown in Kansl2 overexpressing MuSCs. Reference gene: m36b4 (** p = 0.0026, **** p < 0.0001, n = 3). l Quantification of micronuclei-containing cells in scrambled control, Nf-ya knockdown, and Nf-ya knockdown Kansl2 overexpressing MuSCs (**** p < 0.0001, n = 4) from 8-20 weeks old males and females, isolated 4–6 weeks after tamoxifen administration. Data are mean ± SEM of biological replicates. ( a , b , d , f , k ) unpaired two-tailed t test. ( g , h , l ) one-way ANOVA with Bonferroni’s multiple comparisons test. ( i ) one-way ANOVA with Tukey’s multiple comparisons test. ( j ) one-way ANOVA with Holm–Sidak’s multiple comparisons test. Source data are provided in the Source Data file.

Article Snippet: In brief, 1 × 10 6 wild type in vitro cultured muscle stem cells (MuSCs) or 4 × 10 5 wild type and Taf4a sKO MuSCs were immobilized on Concanavalin A-coated magnetic beads (Bangs Laboratories), permeated with 0.05% Digitonin (EMD Millipore), and incubated with TAF4A (Santa Cruz #sc-136093), KANSL2 (Proteintech #27261-1-AP), NF-YA (Santa Cruz #sc-17753) or mouse IgG (Millipore #12-371B) or rabbit IgG (Diagenode #C15410206) on a rotator with 1:100 dilution at 4 °C overnight on a rotator.

Techniques: Staining, Immunofluorescence, Cell Culture, Control, Western Blot, Immunoprecipitation, Quantitative RT-PCR, Expressing, Knockdown, Isolation, Two Tailed Test

Journal: Nature Communications

Article Title: Regulation of NSL by TAF4A is critical for genome stability and quiescence of muscle stem cells

doi: 10.1038/s41467-025-64402-1

Figure Lengend Snippet: TAF4A drives expression of Kansl2 by interacting with the NF-Y complex. KANSL2 is a critical part of the NSL (non-specific lethal) complex, which also comprises the acetyl-transferase MOF. Decreased expression of Kansl2 prevents NSL-mediated acetylation of Lamin, increasing Lamin phosphorylation and solubility. Created in BioRender. Guo, X. (2025) https://BioRender.com/frmva1q .

Article Snippet: In brief, 1 × 10 6 wild type in vitro cultured muscle stem cells (MuSCs) or 4 × 10 5 wild type and Taf4a sKO MuSCs were immobilized on Concanavalin A-coated magnetic beads (Bangs Laboratories), permeated with 0.05% Digitonin (EMD Millipore), and incubated with TAF4A (Santa Cruz #sc-136093), KANSL2 (Proteintech #27261-1-AP), NF-YA (Santa Cruz #sc-17753) or mouse IgG (Millipore #12-371B) or rabbit IgG (Diagenode #C15410206) on a rotator with 1:100 dilution at 4 °C overnight on a rotator.

Techniques: Expressing, Phospho-proteomics, Solubility