Journal: bioRxiv

Article Title: CEBPβ regulates myoblast proliferation and myogenic differentiation during human myogenesis and rescues defective differentiation in FSHD

doi: 10.1101/2025.08.07.669099

Figure Lengend Snippet: (A) Schematic of CEBPB mRNA transcript and protein isoform generation. Translation initiation at three alternative start codons along the CEBPB mRNA transcript yields distinct protein isoforms: LAP* (44 kDa), LAP (38 kDa) and LIP (20 kDa). The two larger isoforms, LAP* and LAP, contain all three protein domains, while the truncated LIP isoform lacks the trans-activation domain (orange) and part of the regulatory domain (pink). (B) CEBPB mRNA expression levels in publicly available RNA-sequencing data from two human myogenic cell lines cultured as myoblasts (MB) or differentiated into myotubes (MT). Differential expression analysis was performed using DESeq2 with the Wald test and Benjamini-Hochberg correction for multiple testing, where ns denotes p > 0.05 and an asterisk denotes p < 0.05. (C) Schema and representative brightfield images from each stage of in vitro myogenic differentiation examined. Scale bars represent 100 μm. Western blot analysis of CEBPB expression through human myogenesis, with protein levels assessed in myoblasts (MB), myocytes (MC) and myotubes (MT) differentiated from control 54.6 and 16U independent myogenic cell lines. Vinculin (VCL) was used as loading control. (D) Bar plots display quantification of Western blots from (C). Relative density unit (RDU) for LAP*+LAP protein isoform expression normalized to Vinculin (VCL) (top). N= 6 replicates collected over two independent experiments. One-way ANOVA + Tukey post-hoc test, where a single asterisk denotes p < 0.05, two asterisks denotes p < 0.01, and four asterisks denotes p < 0.0001, and ns denotes p > 0.05. (E) Stacked bar plots display isoform ratios of LAP*+LAP vs. LIP calculated relative to total CEBPB expression. N= 6 replicates collected over two independent experiments. One-way ANOVA + Tukey post-hoc test, where ns denotes p > 0.05.

Article Snippet: Respective membranes were incubated overnight at 4°C in either TBS-T + 5% milk + 1:10000 anti-vinculin primary antibody (Sigma, #V9131) or TBS-T + 5% BSA + 1:1000 anti-CEBPB primary antibody (Cell Signaling Technology, #90081), followed by secondary antibody incubation for 1 hr at room temperature (RT) in TBS-T + 5% milk + 1:5000 anti-rabbit-HRP (Sigma, #GENA934) or 1:5000 anti-mouse-HRP (Sigma, #NA931V) secondary antibodies.

Techniques: Activation Assay, Expressing, RNA Sequencing, Cell Culture, Quantitative Proteomics, In Vitro, Western Blot, Control

Journal: bioRxiv

Article Title: CEBPβ regulates myoblast proliferation and myogenic differentiation during human myogenesis and rescues defective differentiation in FSHD

doi: 10.1101/2025.08.07.669099

Figure Lengend Snippet: (A) 54.6-iCEBPB and 16U-iCEBPB human myoblast lines were genetically modified to over-express CEBPB (CEBPB O.E.) after exposure to doxycycline (DOX) and were cultured in GM or GM containing DOX for 48 hrs, at which time a 2 hr EdU pulse was performed. Unmodified 54.6 and 16U human myoblast lines were also cultured in GM containing either negative control siRNA (siNC) or anti-CEBPB siRNA (siCβ) overnight, after which growth medium was replaced and EdU pulse performed the following day (2 days post-transfection). (B) Western blots display representative increased CEBPB levels in 54.6-iCEBPB and 16U-iCEBPB cell lines cultured in presence of doxycycline (DOX) for 24 hours (left), and down-regulation of CEBPB in unmodified control 54.6 and 16U myoblasts 48 hours post-transfection with anti-CEBPB (siCβ) siRNA (right), compared to negative control (siNC). Vinculin (VCL) was used as a loading control. (C) Over-expression of CEBPB reduces the myoblast proliferation rate. Quantification of EdU incorporation in 54.6-iCEBPB and 16U-iCEBPB myoblasts following 48 hrs of doxycycline treatment (50 ng/mL). Representative images of incorporated EdU with a Hoechst nuclear counterstain. Bar plots display the percentage of EdU+ nuclei per sample and the total number of nuclei per FOV per sample relative to controls. N=8 replicates per cell line collected over two independent experiments, and each data point represents the average of 3 FOVs per replicate. Total number of nuclei was quantified per sample and divided by the control average of its experiment. Significance was assessed by unpaired t-test, where a single asterisk denotes p < 0.05, two asterisks denotes p < 0.01, four asterisks denotes p < 0.0001, and ns denotes p > 0.05. Scale bars represent 200 μm. (D) Decrease in myoblast proliferation rate following knock-down of CEBPB. Quantification of EdU incorporation in control 54.6 and 16U myoblasts 48 hrs post-transfection with either negative control (siNC) or anti-CEBPB (siCβ) siRNA. Representative images of incorporated EdU with a Hoechst nuclear counterstain. Bar plots display the percentage of EdU+ nuclei per sample and the total number of nuclei per FOV per sample relative to controls. N=8 replicates per cell line collected over two independent experiments, and each data point represents the average of 3 FOVs per replicate. Total number of nuclei per sample was divided by the control average of its experiment. Significance was assessed by unpaired t-test, where a single asterisk denotes p < 0.05, two asterisks denotes p < 0.01, four asterisks denotes p < 0.0001, and ns denotes p > 0.05. Scale bars represent 200 μm.

Article Snippet: Respective membranes were incubated overnight at 4°C in either TBS-T + 5% milk + 1:10000 anti-vinculin primary antibody (Sigma, #V9131) or TBS-T + 5% BSA + 1:1000 anti-CEBPB primary antibody (Cell Signaling Technology, #90081), followed by secondary antibody incubation for 1 hr at room temperature (RT) in TBS-T + 5% milk + 1:5000 anti-rabbit-HRP (Sigma, #GENA934) or 1:5000 anti-mouse-HRP (Sigma, #NA931V) secondary antibodies.

Techniques: Genetically Modified, Cell Culture, Negative Control, Transfection, Western Blot, Control, Over Expression, Knockdown

Journal: bioRxiv

Article Title: CEBPβ regulates myoblast proliferation and myogenic differentiation during human myogenesis and rescues defective differentiation in FSHD

doi: 10.1101/2025.08.07.669099

Figure Lengend Snippet: (A) Box plot displays expression of CEBPB in bulk transcriptomic data from muscle biopsies from 6 healthy control individuals (light blue) and 7 patients with DMD (dark blue) reported by Khairallah et al. . Normalized counts were downloaded from the original publication’s supplementary data . Boxes represent the interquartile range (IQR), lines display the median, and whiskers depict the smallest and largest values within 1.5*IQR from the IQR. Statistical significance was assessed by Wilcoxon ranked-sum test, where ns denotes p > 0.05. Box plot displays expression of CEBPB in bulk transcriptomic data from muscle biopsies from 10 healthy control individuals (light green) and 36 patients with DM1 (dark green) reported by Wang et al. . Normalized counts were downloaded from the original publication’s supplementary data . Boxes represent the interquartile range (IQR), lines display the median, and whiskers depict the smallest and largest values within 1.5*IQR from the IQR. Statistical significance was assessed by Wilcoxon ranked-sum test, where ns denotes p > 0.05. (B) Violin plots display expression of CEBPB in Banerji et al. 2023 and Wong et al. datasets. Differential expression analysis was performed using DESeq2 with the Wald test and Benjamini-Hochberg correction for multiple testing, where a single asterisk denotes p > 0.05, two asterisks denotes p < 0.01, and four asterisks denotes p < 0.0001. All analyses were unpaired, with the exception of TIRM − vs. TIRM + samples in Banerji dataset, in which a paired analysis was performed between matched samples from individual patients. Boxes represent the interquartile range (IQR), lines display the median, and whiskers depict the smallest and largest values within 1.5*IQR from the IQR. (C) Expression of CEBPB in RNA-sequencing data from 54.6 control cell line (blue) and 54.12 FSHD myoblast cell line (red), as well as 16U control cell line (blue) and 16A FSHD cell line (red) at various time points throughout in vitro myogenesis . The ‘54s’ cell line pair was isolated from a mosaic patient, with 54.6 (blue) derived from a clone containing a healthy genetic background and 54.12 (red) containing the disease-conferring contraction of the D4Z4 region. The ‘16s’ cell lines were derived from an FSHD patient (‘16A’, red) and a sibling matched control (‘16U’, blue). (D) Decrease in CEBPB expression in response to endogenous and over-expressed DUX4, which begins as soon as 4 hours after DUX4 induction. Log fold-changes and RNA-sequencing data, along with their corresponding statistics, were obtained from the Supplementary materials from Jagannathan et al. or Campbell et al. . (F, G) CEBPB protein expression was compared in two pairs of control vs. FSHD immortalized myoblast cell lines. Each pair was differentiated in parallel, and protein samples were collected from proliferating myoblasts (MB), from differentiating myocytes (MC) 24 hours after induction of differentiation, and from myotubes (MT). CEBPB protein expression was evaluated by Western blot, and levels of LAP*/LAP isoforms were quantified relative to vinculin (VCL). Colorimetric image of pre-stained protein ladder shown on right to estimate molecular weights. N=6 replicates per cell line collected from two independent experiments. Statistical significance was assessed by unpaired t-test, with one asterisk denoting p < 0.05, three asterisks denoting p < 0.001, and ns denoting p > 0.05.

Article Snippet: Respective membranes were incubated overnight at 4°C in either TBS-T + 5% milk + 1:10000 anti-vinculin primary antibody (Sigma, #V9131) or TBS-T + 5% BSA + 1:1000 anti-CEBPB primary antibody (Cell Signaling Technology, #90081), followed by secondary antibody incubation for 1 hr at room temperature (RT) in TBS-T + 5% milk + 1:5000 anti-rabbit-HRP (Sigma, #GENA934) or 1:5000 anti-mouse-HRP (Sigma, #NA931V) secondary antibodies.

Techniques: Expressing, Control, Quantitative Proteomics, RNA Sequencing, In Vitro, Isolation, Derivative Assay, Western Blot, Staining

Journal: The FASEB Journal

Article Title: Maternal Fructose Intake During Pregnancy Induced the Hepatic Glucose Homeostasis Imbalance in the Offspring by Inhibiting Glucokinase

doi: 10.1096/fj.202503081R

Figure Lengend Snippet: C/EBPβ promoted DNMT3B expression through transcriptional activation. (A) The results of PROMO database prediction. (B, C) Effects of maternal fructose intake during pregnancy on the C/EBPβ expression in the liver of offspring at 0 days and 4 weeks. Actin was used as a loading control ( n = 4 offspring samples per group). (D) Knockdown of C/EBPβ inhibited DNMT3B expression at the protein level in AML12 cells. Actin was used as a loading control ( n = 3 samples per group). (E) Dual‐Luciferase Assay revealed that C/EBPβ trans‐activates the Dnmt3b promoter ( n = 6 samples per group). (F) CHIP‐PCR results showed that C/EBPβ bound to the Dnmt3b promoter. All data were presented as mean ± SEM. * p < 0.05, ** p < 0.01, and **** p < 0.0001.

Article Snippet: The primary antibodies used were as follows: GK (Abclonal, A15059, 1:1000), DNMT3B (Abclonal, A22658, 1:5000), HK2 (Proteintech, 22029‐1‐AP, 1:1500), GLUT2 (Proteintech, 20436‐1‐AP, 1:1500), KHK (Proteintech, 15681‐1‐AP, 1:1000), ALDOB (Proteintech, 18065‐1‐AP, 1:5000), ACLY (Proteintech, 15421‐1‐AP, 1:1000), Actin (Bioworld, BS6007M, 1:5000), and C/EBPβ (Proteintech, 23 431‐1‐AP, 1:15,000).

Techniques: Expressing, Activation Assay, Control, Knockdown, Luciferase

Journal: The FASEB Journal

Article Title: Maternal Fructose Intake During Pregnancy Induced the Hepatic Glucose Homeostasis Imbalance in the Offspring by Inhibiting Glucokinase

doi: 10.1096/fj.202503081R

Figure Lengend Snippet: Schematic diagram of the core mechanisms. The study demonstrated that maternal fructose intake during pregnancy led to glucose intolerance and insulin resistance in offspring. The abnormal hepatic glucose metabolism in offspring was associated with decreased GK expression, which was inhibited by DNMT3B. Besides, the expression of DNMT3B was regulated by C/EBPβ at the transcriptional level. This condition was improved by the administration of dorzagliatin.

Article Snippet: The primary antibodies used were as follows: GK (Abclonal, A15059, 1:1000), DNMT3B (Abclonal, A22658, 1:5000), HK2 (Proteintech, 22029‐1‐AP, 1:1500), GLUT2 (Proteintech, 20436‐1‐AP, 1:1500), KHK (Proteintech, 15681‐1‐AP, 1:1000), ALDOB (Proteintech, 18065‐1‐AP, 1:5000), ACLY (Proteintech, 15421‐1‐AP, 1:1000), Actin (Bioworld, BS6007M, 1:5000), and C/EBPβ (Proteintech, 23 431‐1‐AP, 1:15,000).

Techniques: Expressing

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Super enhancer-driven LINC01013 mediates hypoxia-induced mitochondrial dysfunction by HSPA9 to determine pulmonary arterial smooth muscle cell fate

doi: 10.1007/s00018-025-06071-3

Figure Lengend Snippet: SE-associated LINC01013 was transcriptionally activated by CEBPB in hPASMCs. A Prediction of candidate transcription factors and binding sites. (Transcription factor related databases: JASPAR: https://jaspar.elixir.no/ ; PROMO: https://alggen.lsi.upc.es/cgi-bin/promo_v3/promo/promoinit.cgi?dirDB=TF_8.3 ; GENECARD: https://www.genecards.org/ ; AnimalTFDB: http://bioinfo.life.hust.edu.cn/HumanTFDB/#!/ ; Super enhancer related database: LncSEA: https://bio.liclab.net/LncSEA/ ). B The schematic diagram illustrates the LINC01013 promoter, divided into segments P1-P4, and the binding sites of transcription factors. C Four constituents (E1-E4) of SE region of LINC01013 derived from the WashU Epigenome Browser databases ( http://epigenomegateway.wustl.edu/browser/ ). D , E hPASMCs were subjected to ChIP analysis using antibodies against H3K27ac, H3K4me1 and CEBPB. The association with the SE region (D, E1-E4) and promoter region (E, P1-P4) of LINC01013 was quantified by RT‒qPCR ( n = 3). F hPASMCs were treated with CEBPB siRNA and subjected to ChIP analysis using antibodies against H3K27ac. The association with the E2 of SE (left) and P1-P3 promoter regions (right) of LINC01013 was quantified by RT-qPCR ( n = 3). G Schematic diagram of transcribing LINC01013 in hPASMCs. All values are presented as the mean ± SEM. Statistical analysis was performed with one-way ANOVA or Student’s t-test. * p < 0.05, ** p < 0.01, *** p < 0.001. Nor, normoxia; Hyp, hypoxia; NC, negative control; IP, immunoprecipitation; IgG, Immunoglobulin G; TSS, transcription initiation site

Article Snippet: The antibody against CEBPB (PB9171, BA0670, 1:500, Boster, Wuhan, China), H3K27ac (A7253, 1:500, ABclonal, Wuhan, China), H3K4me1 (A2355, 1:500, Wuhan, China), PCNA (A00125, 1:500, Boster, Wuhan, China), Cyclin A (PB0515, 1:500, Boster, Wuhan, China), Cyclin D (BM4272, 1:500, Boster, Wuhan, China), IL-6 (AF7236, 1:500, Beyotime, Shanghai, China), TNF-α (AF8208, 1:500, Beyotime, Shanghai, China), PKM2 (4053, 1:1000, Cell Signaling, MA, US), HK II (66974-1-Ig, 1:1000, Proteintech, IL, USA), PDH (2784, 1:1000, Cell Signaling, MA, US), HSPA9 (14887-1-AP, 1:5000, Proteintech, IL, USA), VDAC1 (10866-1-AP, 1:5000, Proteintech, IL, USA), and β-actin (TA-09, 1:1000, ZSGB‐BIO, Beijing, China) was incubated at 4 °C overnight, followed by incubation with appropriate horseradish peroxidase-conjugated secondary antibodies at room temperature for 1 h, and proteins were visualized with enhanced chemiluminescence reagents.

Techniques: Binding Assay, Derivative Assay, Quantitative RT-PCR, Negative Control, Immunoprecipitation

Journal: Physiological Reports

Article Title: Effects of preconditioning with heat stress on acute exercise‐induced intracellular signaling in male rat gastrocnemius muscle

doi: 10.14814/phy2.15913

Figure Lengend Snippet: Correlations between heat shock protein 72 level and signaling level in both red and white regions of the gastrocnemius muscle.

Article Snippet: The membranes were blocked in a blocking buffer (EveryBlot; Bio‐Rad) for 20 min. After several washes, the membranes were incubated overnight with primary antibodies against the following proteins: phosphorylated Ser473‐Akt (1:2000; catalog #4060), phosphorylated Thr308‐Akt (1:2000; #2965), Akt (1:2000; #2920), phosphorylated Ser2448‐mTOR (1:2000; #2971), mTOR (1:2000; Cell Signaling, #2983), phosphorylated Thr389‐p70S6 kinase (1:2000; #9234), p70S6K (1:2000; #9202), phosphorylated Thr37/46‐4E‐BP1 (1:2000; #2855), and 4E‐BP1 (1:2000; #9452), phosphorylated Thr202/Tyr204 ERK1/2 (1:2000; #4370), ERK (1:2000; #4695), phosphorylated Thr180/Tyr182 p38 MAPK (1:2000; #4511), p38 MAPK (1:2000; #8690), phosphorylated Ser253‐FoxO3a (1:2000; #13129), c‐Myc (1:2000; #9402), and LC3A/B (1:2000; #4108) (all from Cell Signaling Technology, Beverly, MA, USA); FoxO3a (1:2000; 04‐1007) and Puromycin (1:2000; MABE343) (from Millipore, Temecula, CA, USA); and 4‐hydroxynonenal (4‐HNE) (1:2000; ab46545, Abcam, Cambridge, MA, USA).

Techniques: Phospho-proteomics

Journal: Physiological Reports

Article Title: Effects of preconditioning with heat stress on acute exercise‐induced intracellular signaling in male rat gastrocnemius muscle

doi: 10.14814/phy2.15913

Figure Lengend Snippet: Correlations between the heat shock protein 72 and signaling molecules, as well as their increased levels, in each CT leg in the red and white regions of the gastrocnemius muscle following downhill exercise.

Article Snippet: The membranes were blocked in a blocking buffer (EveryBlot; Bio‐Rad) for 20 min. After several washes, the membranes were incubated overnight with primary antibodies against the following proteins: phosphorylated Ser473‐Akt (1:2000; catalog #4060), phosphorylated Thr308‐Akt (1:2000; #2965), Akt (1:2000; #2920), phosphorylated Ser2448‐mTOR (1:2000; #2971), mTOR (1:2000; Cell Signaling, #2983), phosphorylated Thr389‐p70S6 kinase (1:2000; #9234), p70S6K (1:2000; #9202), phosphorylated Thr37/46‐4E‐BP1 (1:2000; #2855), and 4E‐BP1 (1:2000; #9452), phosphorylated Thr202/Tyr204 ERK1/2 (1:2000; #4370), ERK (1:2000; #4695), phosphorylated Thr180/Tyr182 p38 MAPK (1:2000; #4511), p38 MAPK (1:2000; #8690), phosphorylated Ser253‐FoxO3a (1:2000; #13129), c‐Myc (1:2000; #9402), and LC3A/B (1:2000; #4108) (all from Cell Signaling Technology, Beverly, MA, USA); FoxO3a (1:2000; 04‐1007) and Puromycin (1:2000; MABE343) (from Millipore, Temecula, CA, USA); and 4‐hydroxynonenal (4‐HNE) (1:2000; ab46545, Abcam, Cambridge, MA, USA).

Techniques: Phospho-proteomics

Journal: The Journal of cell biology

Article Title: BAR scaffolds drive membrane fission by crowding disordered domains.

doi: 10.1083/jcb.201807119

Figure Lengend Snippet: Figure 1. Amphiphysin drives membrane fission, while the N-BAR domain stabilizes membrane tubules. Mem- brane composition for vesicles in TEM: 80 mol% DOPC, 5 mol% PtdIns(4,5)P2, and 15 mol% DOPS. SUPER template membrane composition: 79 mol% DOPC, 5 mol% PtdIns(4,5)P2, 15 mol% DOPS, and 1 mol% Texas Red–DHPE. (A) Schematic of Amph-FL dimer. BAR domain: PDB 4ATM. SH3 domain: PDB 1BB9. (B–D) Negative stain TEM micrographs of 200 nm extruded vesicles before exposure to protein (B), after exposure to 26 µM N-BAR (C), and after exposure to 5 µM Amph-FL (D). Dashed boxes indicate zoomed regions to the right. Black arrows indi- cate membrane tubules; red arrowheads indicate fission vesi- cles. Yellow asterisks indicate small vesicles that are present in the vesicle population before protein exposure. (E) Histograms of vesicle diameters measured from electron micrographs. Ves- icles alone: n = 1,302 vesicles. 26 µM N-BAR: n = 462 vesicles. 5 µM Amph-FL: n = 1,071 vesicles. (F) Membrane release from SUPER templates, measured as Texas Red signal present in the supernatant after sedimentation of the SUPER templates. Membrane release in the absence of protein was measured and subtracted as background. Dots indicate data and lines indi- cate mean; n = 3 independent experiments. P value: one-tailed, unpaired Student’s t test. (B–D) Bars, 500 nm; insets, 200 nm. See also Fig. S1 and Videos 1, 2, 3, 4, and 5.

Article Snippet: The pCAG EN mammalian expression vector containing the N-BAR domain of human amphiphysin (residues 1–256), tagged at the C terminus with mCherry, was a gift from T. Meyer, Stanford University, Stanford, CA (Addgene plasmid 85130).

Techniques: Membrane, Staining, Sedimentation, One-tailed Test

Journal: The Journal of cell biology

Article Title: BAR scaffolds drive membrane fission by crowding disordered domains.

doi: 10.1083/jcb.201807119

Figure Lengend Snippet: Figure 3. The disordered domain of amphiphysin alone drives membrane fission, but the N-BAR scaffold substantially enhances fission efficiency. Membrane composition in Amph CTD ΔSH3 tethered vesicle experiments: 76 mol% DOPC, 20 mol% DOGS- NTA-Ni, 2 mol% Oregon Green 488–DHPE, and 2 mol% DP-EG10-biotin. In tethered vesicle experiments with N-BAR-epsin CTD, DOGS-NTA-Ni was replaced with 5 mol% PtdIns(4,5)P2 and 15 mol% DOPS. SUPER template membrane composition: 79 mol% DOPC, 5 mol% PtdIns(4,5)P2, 15 mol% DOPS, and 1 mol% Texas Red–DHPE. (A) Schematic of Amph CTD ΔSH3. (B) Tethered vesicle fission experiments show that Amph CTD ΔSH3 forms highly curved fission products. (C) Summary of data from tethered vesicle fission exper- iments with Amph CTD ΔSH3 expressed as the ratio of the distribution area below 45 nm to the total distri- bution area (compare to Fig. 2 F). (D) Coverage of the membrane surface by Amph CTD ΔSH3 and Amph-FL as a function of protein concentration. Amph-FL data from Fig. 2 H. (E) Fraction of vesicle diameters below 45 nm generated by Amph CTD ΔSH3 and Amph-FL versus coverage of the membrane surface by proteins. Amph-FL fission data from Figs. 2 F and S2 M. Amph CTD ΔSH3 fission data from Fig. 3 C. (F) Schematic of N-BAR-epsin CTD chimera dimer. (G) Tethered vesicle fission measurements show that N-BAR-epsin CTD generates highly curved fission vesicle populations over the concentration range of 10–150 nM, similar to Amph-FL (compare to Fig. 2 D). (H) Summary of data from tethered vesicle fission experiments with N-BAR-epsin CTD, expressed as the ratio of the dis- tribution area below 45 nm to the total distribution area. Amph-FL and N-BAR data from Fig. 2 F. (I) SUP ER template membrane shedding experiments show that N-BAR-epsin CTD drives greater membrane release compared with N-BAR (compare to Fig. 1 F). Dots indicate data and lines indicate mean; n = 3 inde- pendent experiments. P value: one-tailed, unpaired Student’s t test. Amph CTD ΔSH3 markers in C and D and all markers in H represent mean ± first SD; n = 3 independent experiments. (J) Schematic of the N-BAR scaffold (EMDB 3192; Adam et al., 2015) with attachment points of some of the disordered domains marked (two per N-BAR dimer). Dashed circles indi- cate approximate volumes occupied by undeformed disordered domains. See also Figs. S2 and S3.

Article Snippet: The pCAG EN mammalian expression vector containing the N-BAR domain of human amphiphysin (residues 1–256), tagged at the C terminus with mCherry, was a gift from T. Meyer, Stanford University, Stanford, CA (Addgene plasmid 85130).

Techniques: Membrane, Protein Concentration, Generated, Concentration Assay, One-tailed Test