An effective NIR fluorescent molecular tool to monitor cancer cell migration derived from inhibiting autophagy-induced cellular inflammation

Yang, Jialu; Duan, Lizheng; Zhou, Yunhao; Wu, Tian; Shi, Jiaoying; Zhou, Yanmei

doi:10.1016/j.dyepig.2022.110846

Cited by 4 publications

(2 citation statements)

References 32 publications

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“…Different from the mechanism of starvation-induced autophagy, rapamycin can induce autophagic flux through its ability to inhibit the activity of mammalian target of rapamycin (mTOR). 19,20,36,37 Using fluorescence imaging techniques and FCS (Fig. S13A and S13B†), we observed that in the presence of CQ, H1299 cells treated with rapamycin had more and more EGFP-LC3B punctae with increasing incubation time, and reached the maximum value (about 19.3%) between 2 and 3 h. Therefore, we investigated rapamycin-induced autophagy flux at 2 h in this study.…”

Section: Resultsmentioning

confidence: 99%

In vivo measurement of autophagic flux by fluorescence correlation spectroscopy

Song

Dong

Ren

2023

Analyst

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Section: Resultsmentioning

confidence: 99%

In vivo measurement of autophagic flux by fluorescence correlation spectroscopy

Song

Dong

Ren

2023

Analyst

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“…Flow cytometry, as a high-throughput fluorescence method, can rapidly assess autophagic flux in living cells. , However, its incapability to monitor autophagic structures limits its study of macroautophagy . Fluorescence imaging technique is a direct method to visualize the change of autophagosomes in living cells. − This method is able to distinguish autophagosomes and autolysosomes by simultaneously labeling with two fluorescent proteins of different acid sensitivities . Thus, the autophagosome production and delivery to lysosome can be monitored by this method .…”

Section: Introductionmentioning

confidence: 99%

Simultaneously Monitoring Multiple Autophagic Processes and Assessing Autophagic Flux in Single Cells by In Situ Fluorescence Cross-Correlation Spectroscopy

Song,

Dong,

Ren

2024

Anal. Chem.

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Autophagy is a widely conserved and multistep cellular catabolic process and maintains cellular homeostasis and normal cellular functions via the degradation of some harmful intracellular components. It was reported that high basal autophagic activity may be closely related to tumorigenesis. So far, the fluorescence imaging technique has been widely used to study autophagic processes, but this method is only suitable for distinguishing autophagosomes and autolysosomes. Simultaneously monitoring multiple autophagic processes remains a significant challenge due to the lack of an efficient detection method. Here, we demonstrated a new method for simultaneously monitoring multiple autophagic processes and assessing autophagic flux in single cells based on in situ fluorescence cross-correlation spectroscopy (FCCS). In this study, microtubule-associated protein 1A/1B-light chain 3B (LC3B) was fused with two tandem fluorescent proteins [mCherry red fluorescent protein (mCherry) and enhanced green fluorescent protein (EGFP)] to achieve the simultaneous labeling and distinguishing of multiple autophagic structures based on the differences in characteristic diffusion time (τD). Furthermore, we proposed a new parameter “delivery efficiency of autophagosome (DE AP)” to assess autophagic flux based on the cross correlation (CC) value. Our results demonstrate that FCCS can efficiently distinguish three autophagic structures, assess the induced autophagic flux, and discriminate different autophagy regulators. Compared with the commonly used fluorescence imaging technique, the resolution of FCCS remains unaffected by Brownian motion and fluorescent monomers in the cytoplasm and is well suitable to distinguishing differently colored autophagic structures and monitoring autophagy.

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