Part 1A: Confocal Microscopy (Brain Slice 1) - Raw Data for MS: 10.1016/j.nbd.2023.106277; Title" 20 kDa isoform of connexin-43 augments reorganization of the junctional in CCM3

Description

NOTICE TO AUDITORS: This dataset serves as Folder 1A and the master index for an interconnected, 8-part 53.8GB raw evidentiary repository hosted natively within Elsevier via Mendeley Data. The home institution's inquiry committee and the journal's editorial office explicitly declined to review or independently audit these raw files during the appeal process. They are published openly here for transparent, independent global peer review to protect scientific truth and ensure due process. DATA COMPARTMENTALIZATION ROADMAP: • Confocal Microscopy Data: Segmented across Folders 1A, 1B, 1C, 2A, and 2B. • Western Blot Data Replicates: Segmented across Folders 3, 4, 5, 6, 7a, and 7b. Please use the "Related Links" sidebar to navigate seamlessly between these compartments. 1. CONFOCAL MICROSCOPY DATA AUDIT LEDGER Original microscopy materials associated with concerns in Figures 2E and 6I (unedited Z-stacks, MIP files, optical sections, and orthogonal/3D views): • 1.1 Folder 1A (Here): Brain Slice 1 - Leaky lesion (LL). Markers: Anti-ZO-1 / anti-claudin-5. • 1.2 Folder 1B: Brain Slice 2 - Leaky lesion (LL). Markers: Anti-ZO-1 / N-terminal Cx43. • 1.3 Folder 1C: Brain Slice 3 - Leaky & non-leaky regions (LL + NL). Markers: Anti-ZO-1 / anti-claudin-5. Includes additional slice 3CCM3KOC-CX43N-CX43 ZO-1 LL. • 1.4 Folder 2A: Brain Slice 4 - Non-leaky region (NL). Markers: Anti-ZO-1 / C-terminal Cx43. Includes additional slice 2CCM3KOcldn5Zo-1Cx43 NLL. • 1.5 Folder 2B (Control Slice 1): Markers: Anti-ZO-1 / anti-claudin-5. • 1.6 Folder 2B (Control Slice 2): Markers: Anti-ZO-1 / Cx43. 2. WESTERN BLOT REPLICATES AUDIT LEDGER Complete uncropped digital captures and raw instrument films confirming the 20 kDa Cx43 isoform: • 2.1 Folder 3 (Upstream MNK/mTOR Screen): Lane order verified via student record "110518 meeting.pptx" (Slides 1-4). Lanes 1-2 are controls; lanes 5-6 are CCM3KD replicates. • 2.2 Folder 4 (Electroporation Pathway Validation): Multi-experiment validation cohorts (Exp 1, 2, and 3). Keys provided via associated .docx/.pdf sheets. Quantifications mapped via .xlsx and .pzfx layouts. First two lanes serve as original unedited replicates suitable for Figure 3A replacement. • 2.3 Folder 5 (Downstream Signaling): Exposure matrices (1s-60s) contrasting commercial antibody performance profiles (Abcam vs. CST). Loading lane maps tracked natively via master key Cx43KD.xlsx. • 2.4 Folder 6 (Primary Cells & ERK Inhibition): Raw data evaluating ERK1/2 pathway inhibition parameters (supporting backing data for Manuscript Figure 7). Key provided via ERK inhibition.xlsx / .pzfx. Contains three independent biological replicates analyzed with N-terminal Cx43 antibody. • 2.5 Folders 7a, 7b and 7b part2 (Primary Endothelial Transfection Optimization): Establishes technical control and protocol delivery baselines. This dataset stores unedited raw instrument files in its native formats, preserved from the source computer for transparency and tracking.

Steps to reproduce

EXPERIMENTAL VERIFICATION METHODOLOGY: 1. CONFOCAL MICROSCOPY DATA CORRECTION: To verify that overlapping structures in Folders 1A, 1B, 1C, and 2A are benign physical compression artifacts rather than manual digital duplication, auditors must download the raw multi-channel files. Open the proprietary files (.czi/.lif/.oib) using open-source imaging software (Fiji / ImageJ). Scroll frame-by-frame along the Z-axis (focal depth). Reviewers will confirm that distinct cellular features disappear and reappear at structurally independent Z-depth horizons, and that distinct frames possess unique metadata and camera timestamps. 2. WESTERN BLOT AUDIT COMPLIANCE: To verify the existence and reproducibility of the 20 kDa Cx43 isoform, reviewers are directed to Folders 3 through 7b. Open the uncut, full-frame digital film captures (.tif). Cross-reference the lane layouts with the corresponding spreadsheet keys (.xlsx) or student tracking presentation decks (.pptx) enclosed within each respective folder. The presence of flanking protein molecular ladders on the unedited gel margins rules out any selective lane-cropping, band-splicing, or intentional data adjustment.

Institutions

• University of Michigan

  Michigan, Ann Arbor

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