Journal: bioRxiv

Article Title: c-MYC is an aggregation-prone, amyloidogenic protein

doi: 10.64898/2026.03.12.711438

Figure Lengend Snippet: (A) Detection of detergent-soluble and - insoluble c-MYC proteins by immunoblotting in HeLa cells pre-treated with 30 μM CR, followed by HS at 43°C for 30 min (representative images of three independent experiments). (B) and (C) Immunoprecipitation of either exogenously expressed HA-c-MYC or endogenous c-MYC proteins in HeLa cells by A11 antibodies (representative images of three independent experiments). (D) Detection of endogenous c-MYC proteins in HeLa cells by PLA using both the goat anti-c-MYC Ab and the rabbit anti-AO (A11) Ab. Nuclei are counterstained with Hoechst 33342. Scale bar: 10 μm. (E)-(G) Detection of endogenous c-MYC proteins in human prostate adenocarcinoma, hepatocellular carcinoma, and Alzheimer’s disease brain tissues by brightfield PLA using both the goat anti-c-MYC Ab and the rabbit anti-AO (A11) Ab. Scale bar: 100 μm for low magnification and 10 μm for high magnification. (H): Quantitation of endogenous c-MYC recognized by both the goat anti-c-MYC Ab and the rabbit anti-AO (A11) Ab through In-Cell PLA ELISA in HeLa cells treated with CR (mean ± SD, n=3 independent experiments, One-way ANOVA).

Article Snippet: All antibodies were purchased commercially, including rabbit polyclonal anti-amyloid oligomer (A11) Ab (cat#SPC-506, StressMarq Biosciences Inc.), rabbit monoclonal anti-c-MYC/N-MYC (D3N8F) Ab (cat#13987, Cell Signaling Technology), rabbit monoclonal ani-MAX (E6F6Y) Ab (cat#17471, Cell Signaling Technology), mouse monoclonal anti-HSC/HSP70 (W27) Ab (cat#sc-24, Santa Cruz Biotechnology Inc.), mouse monoclonal anti-HSP90α/β (F-8) Ab (cat#sc-13119, Santa Cruz Biotechnology Inc.), rabbit monoclonal anti-cleaved caspase 3 (Asp175) (D3E9) Ab (cat#9579, Cell Signaling Technology), goat polyclonal anti-c-MYC Ab (cat#AF3696, R&D Systems Inc.), mouse monoclonal anti-HA Ab (cat#901513, BioLegend Inc.), mouse monoclonal anti-βActin (GT5512) Ab (cat#GTX629630, GeneTex Inc.), normal rabbit IgG (cat#02-6102,ThermoFisher Scientific Inc.), Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L) (cat#111-035-144, Jackson ImmunoResearch Inc.), Peroxidase AffiniPure Goat Anti-Mouse IgG (H+L) (cat#115-035-003, Jackson ImmunoResearch Inc.), and Peroxidase IgG Fraction Monoclonal Mouse Anti-Goat IgG, light chain specific (cat#205-032-176, Jackson ImmunoResearch Inc.).

Techniques: Western Blot, Immunoprecipitation, Quantitation Assay, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: c-MYC is an aggregation-prone, amyloidogenic protein

doi: 10.64898/2026.03.12.711438

Figure Lengend Snippet: (A) In vitro fibrillation assays using recombinant c-MYC proteins (mean ± SEM, n=3 independent experiments, Two-way ANOVA). (B) ELISA quantitation of A11 + –AOs generated by recombinant c-MYC proteins in vitro (mean ± SD, n=3 independent experiments, One-way ANOVA). (C) TEM visualization of protofibrils formed by recombinant c-MYC proteins in vitro (representative images of three independent experiments). Scale bars: 100 nm. (D) In vitro fibrillation assays using recombinant c-MYC/MAX dimers (mean ± SEM, n=3 independent experiments, Two-way ANOVA). (E) TEM visualization of recombinant c-MYC/MAX dimers in vitro (representative images of three independent experiments). Scale bars: 100 nm. (F) Quantitation of endogenous c-MYC AOs in HeLa cells treated with 10058-F4 by In-Cell PLA ELISA (mean ± SD, n=3 independent experiments, One-way ANOVA).

Article Snippet: All antibodies were purchased commercially, including rabbit polyclonal anti-amyloid oligomer (A11) Ab (cat#SPC-506, StressMarq Biosciences Inc.), rabbit monoclonal anti-c-MYC/N-MYC (D3N8F) Ab (cat#13987, Cell Signaling Technology), rabbit monoclonal ani-MAX (E6F6Y) Ab (cat#17471, Cell Signaling Technology), mouse monoclonal anti-HSC/HSP70 (W27) Ab (cat#sc-24, Santa Cruz Biotechnology Inc.), mouse monoclonal anti-HSP90α/β (F-8) Ab (cat#sc-13119, Santa Cruz Biotechnology Inc.), rabbit monoclonal anti-cleaved caspase 3 (Asp175) (D3E9) Ab (cat#9579, Cell Signaling Technology), goat polyclonal anti-c-MYC Ab (cat#AF3696, R&D Systems Inc.), mouse monoclonal anti-HA Ab (cat#901513, BioLegend Inc.), mouse monoclonal anti-βActin (GT5512) Ab (cat#GTX629630, GeneTex Inc.), normal rabbit IgG (cat#02-6102,ThermoFisher Scientific Inc.), Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L) (cat#111-035-144, Jackson ImmunoResearch Inc.), Peroxidase AffiniPure Goat Anti-Mouse IgG (H+L) (cat#115-035-003, Jackson ImmunoResearch Inc.), and Peroxidase IgG Fraction Monoclonal Mouse Anti-Goat IgG, light chain specific (cat#205-032-176, Jackson ImmunoResearch Inc.).

Techniques: In Vitro, Recombinant, Enzyme-linked Immunosorbent Assay, Quantitation Assay, Generated

Journal: bioRxiv

Article Title: c-MYC is an aggregation-prone, amyloidogenic protein

doi: 10.64898/2026.03.12.711438

Figure Lengend Snippet: (A) In vitro fibrillation assays using the human c-MYC peptide library (mean ± SEM, n=3 independent experiments, Two-way ANOVA). (B) ELISA quantitation of A11 + –AOs generated by synthetic c-MYC peptides. Results of three independent experiments are presented as a heatmap. FC: fold change. (C) and (D) In vitro fibrillation assays using the P2 and P12 peptides that are subjected to alanine scan. Results are presented as heatmaps using the averages of three independent experiments.

Article Snippet: All antibodies were purchased commercially, including rabbit polyclonal anti-amyloid oligomer (A11) Ab (cat#SPC-506, StressMarq Biosciences Inc.), rabbit monoclonal anti-c-MYC/N-MYC (D3N8F) Ab (cat#13987, Cell Signaling Technology), rabbit monoclonal ani-MAX (E6F6Y) Ab (cat#17471, Cell Signaling Technology), mouse monoclonal anti-HSC/HSP70 (W27) Ab (cat#sc-24, Santa Cruz Biotechnology Inc.), mouse monoclonal anti-HSP90α/β (F-8) Ab (cat#sc-13119, Santa Cruz Biotechnology Inc.), rabbit monoclonal anti-cleaved caspase 3 (Asp175) (D3E9) Ab (cat#9579, Cell Signaling Technology), goat polyclonal anti-c-MYC Ab (cat#AF3696, R&D Systems Inc.), mouse monoclonal anti-HA Ab (cat#901513, BioLegend Inc.), mouse monoclonal anti-βActin (GT5512) Ab (cat#GTX629630, GeneTex Inc.), normal rabbit IgG (cat#02-6102,ThermoFisher Scientific Inc.), Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L) (cat#111-035-144, Jackson ImmunoResearch Inc.), Peroxidase AffiniPure Goat Anti-Mouse IgG (H+L) (cat#115-035-003, Jackson ImmunoResearch Inc.), and Peroxidase IgG Fraction Monoclonal Mouse Anti-Goat IgG, light chain specific (cat#205-032-176, Jackson ImmunoResearch Inc.).

Techniques: In Vitro, Enzyme-linked Immunosorbent Assay, Quantitation Assay, Generated

Journal: Kidney & Blood Pressure Research

Article Title: The Effect of Renal Ischemia/Reperfusion on the Renal Expression of Epithelial Sodium Channels in Rat: Possible Role of Neural Precursor Cell-Expressed Developmentally Down-Regulated Protein (Nedd4-2)

doi: 10.1159/000551003

Figure Lengend Snippet: The effect of ischemia/reperfusion (I/R) on mRNA expression of α-, β-, and γ-ENaC. Sham ( n = 8), IR ( n = 7). Results are expressed as mean ± SEM and compared using unpaired Student’s t test.

Article Snippet: The following antibodies used: polyclonal rabbit anti α-ENaC antibody (#ab214192, Abcam, UK) dilution 1:250, rabbit polyclonal anti-rat β-ENaC (#SPC-404, StressMarq Biosciences Inc., Canada), dilution 1: 100, rabbit polyclonal anti-rat γ-ENaC (#SPC-405, StressMarq Biosciences Inc., Canada) dilution 1:250, rabbit polyclonal anti-Nedd4-2 Antibody (#4013, Cell Signaling Technology, USA0 dilution 1:250, rabbit monoclonal anti-p-Nedd4-2 Phospho S448 antibody (#ab168349, Abcam, UK) dilution 1;250, rabbit monoclonal (Y365) anti-AMPK alpha 1 antibody (#ab32047, Abcam, UK), dilution 1:200 and rabbit recombinant monoclonal anti-AMPK alpha 1 (Phospho T-183) + AMPK alpha 2 (Phospho T172) antibody (EPR5683) (ab133448, Abcam, UK), dilution 1: 150.

Techniques: Expressing

Journal: bioRxiv

Article Title: Differential Assembly of Native ENaC Complexes Across Mouse Epithelial Tissues

doi: 10.64898/2026.01.23.701393

Figure Lengend Snippet: a . Schematic of the tagged ENaCγ subunit produced from the ENaCγ-VF allele, which includes a C-terminal mVenus, 3C protease site, and 3xFLAG tag. b . Representative immunofluorescence image showing co-localization of mVenus and ENaCγ in kidney tissue. mVenus signal was detected using a FITC-conjugated anti-GFP primary antibody (goat, Rockland, 1:100) and Alexa Fluor 488 donkey anti-goat secondary antibody. Endogenous ENaCγ was detected using a rabbit anti-ENaCγ primary antibody (StressMarq, 1:500) targeting the C-terminus, and Alexa Fluor 647 donkey anti-rabbit secondary antibody. DAPI is 4’,6-diamidino-2-phenylindole . c . Close-up view highlighting localization of tagged ENaCγ.

Article Snippet: Endogenous ENaCγ was detected using a rabbit anti-ENaCγ primary antibody (StressMarq, 1:500) targeting the C-terminus, and Alexa Fluor 647 donkey anti-rabbit secondary antibody.

Techniques: Produced, Immunofluorescence

Journal: bioRxiv

Article Title: Differential Assembly of Native ENaC Complexes Across Mouse Epithelial Tissues

doi: 10.64898/2026.01.23.701393

Figure Lengend Snippet: a. Cryo-EM map of human ENaC in complex with two Fabs, 10D4 and 7B1, shown in side and top-down views. In the top-down view, a red box highlights the interaction site between 7B1 and the α subunit. b. Sequence alignment of the experimentally determined 7B1 epitope region in human and mouse ENaCα subunits, revealing high conservation and supporting the potential for cross-reactivity with mouse ENaC. c . Schematic representation of the 7B1-mScarlet Fab construct used to detect the ENaC α subunit. The variable regions of the 7B1 heavy and light chains were expressed as a Fab fragment in HEK293 cells, with mScarlet fused to the C-terminus of the heavy chain. d . FSEC trace monitoring mVenus fluorescence from lung lysates of ENaCγ-VF mice, showing the elution profile of γ-containing ENaC complexes. Dotted lines indicate the elution range for mVenus-tagged complexes (11-16 mL). e . FSEC trace from the same sample following 7B1-mScarlet fluorescence, revealing the elution profile of α-containing ENaC complexes. Dashed lines show that the α-associated signal is restricted to a narrower elution range (11-13 mL), suggesting that α-γ co-assemblies represent a more defined subset of the total γ-containing complexes. f . Overlay of the FSEC traces shown in panels d and e , plotted on the same axes to facilitate direct comparison of elution profiles and peak positions. The inset shows the full normalized traces for each condition. The main panel displays a magnified view of the gray-shaded region indicated in the inset, highlighting the relative alignment of the major peaks.

Article Snippet: Endogenous ENaCγ was detected using a rabbit anti-ENaCγ primary antibody (StressMarq, 1:500) targeting the C-terminus, and Alexa Fluor 647 donkey anti-rabbit secondary antibody.

Techniques: Cryo-EM Sample Prep, Sequencing, Construct, Fluorescence, Comparison