Evidence map›Paper›PMID 40181489›Full record

ArticleFEBS open bio2025

A non-fluorescent immunohistochemistry method for measuring autophagy flux using MAP1LC3/LC3 and SQSTM1 as core markers.

Shahla Shojaei, Amir Barzegar Behrooz, Marco Cordani, Mahmoud Aghaei, Negar Azarpira, Daniel J Klionsky, Saeid Ghavami

Abstract read
In one paragraph

Article in FEBS open bio, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.

0numbers the graph read from it
0cells of the map it votes in
4citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

4 citing papers in PubMed.

  1. Article
  2. Article
  3. Review
  4. Review
4 · The record

Corrections and comments

5 · Who and what money

Authors and funding

7 authors.

Shahla ShojaeiDepartment of Human Anatomy and Cell Science, College of Medicine, University of Manitoba, Winnipeg, Canada.
Amir Barzegar BehroozDepartment of Human Anatomy and Cell Science, College of Medicine, University of Manitoba, Winnipeg, Canada.
Marco CordaniDepartment of Biochemistry and Molecular Biology, Faculty of Biological Sciences, Complutense University, Madrid, Spain.
Mahmoud AghaeiDepartment of Clinical Biochemistry, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran.
Negar AzarpiraShiraz Institute for Stem Cell & Regenerative Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.ORCID https://orcid.org/0000-0002-5549-0057
Daniel J KlionskyLife Sciences Institute and Department of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI, USA.ORCID https://orcid.org/0000-0002-7828-8118
Saeid GhavamiDepartment of Human Anatomy and Cell Science, College of Medicine, University of Manitoba, Winnipeg, Canada.ORCID https://orcid.org/0000-0001-5948-508X

Funding

The mechanism and regulation of autophagyR35GM131919 · NIGMS · UNIVERSITY OF MICHIGAN AT ANN ARBOR · PI DANIEL J. KLIONSKY · 2019 to 2026
$6.4M
CancerCare Manitoba Foundation 763117252NIGMS NIH HHS R35 GM131919NIH HHS GM131919
6 · The paper itself

Abstract

Macroautophagy/autophagy is a crucial cellular process for degrading and recycling damaged proteins and organelles, playing a significant role in diseases such as cancer and neurodegeneration. Evaluating autophagy flux, which tracks autophagosome formation, maturation, and degradation, is essential for understanding disease mechanisms. Current fluorescence-based methods are resource-intensive, requiring advanced equipment and expertise, limiting their use in clinical laboratories. Here, we introduce a non-fluorescent immunohistochemistry (IHC) method using MAP1LC3/LC3 and SQSTM1 as core markers for autophagy flux assessment. LC3 levels reflect autophagosome formation, whereas SQSTM1 degradation and a decrease in the number of its puncta indicate active flux (i.e., lysosomal turnover). We optimized chromogenic detection using diaminobenzidine (DAB) staining and developed a scoring system based on puncta number and the percentage of stained cells. This accessible, cost-effective method enables reliable autophagy quantification using a standard light microscope, bridging the gap between experimental research and clinical diagnostics. Our protocol allows accurate autophagy evaluation in fixed tissues, offering practical applications in biomedical research and clinical pathology assessment.

Indexed as

AutophagyImmunohistochemistryMicrotubule-Associated ProteinsSequestosome-1 ProteinAutophagosomesBiomarkersHumansBiomarkersMAP1LC3A protein, humanMicrotubule-Associated ProteinsSequestosome-1 ProteinSQSTM1 protein, humanautophagometerautophagy flux measurementcellular homeostasis analysischromogenic detectioncost‐effective autophagy assaynon‐fluorescent immunohistochemistry

Identifiers

PMID40181489
PMCPMC12127880

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.