Jan 1, 2024 · Our findings firstly pinpoint BaP exposure as a high-risk factor for SCLC and worse outcomes in patients, with the underlying mechanism being the activation of cancer stemness of SCLC via the AhR/PKA/SOX2 axis. Keywords: Benzo [a]pyrene; Cigarette smoke; PKA; Small cell lung cancer; cancer stemness. Copyright © 2023. Published by Elsevier B.V.

Jan 1, 2024 · Urine 3OH-BaP levels in smokers are systematically evaluated. Low dose BaP exposure upregulates PKA expression and stemness in SCLC. PKA stabilizes SOX2 by counteracting the interaction between SOX2 and COP1. Inhibition of PKA suppresses BaP induced metastasis of SCLC in vitro and in vivo.

Jan 16, 2020 · Here, a detailed protocol for the detection and quantitative analysis of in vivo protein–protein proximity of Sox2 and Oct4 using the proximity-utilizing biotinylation (PUB) method is described.

Treatment with an anti-SOX2 antibody on the third blot indicates recombinant BAP-SOX2 and the absence of a detectable amount of endogenous SOX2. BAP adds a mass shift of 3 kDa, and only a single band was observed on the anti-SOX2 blot in lane 2.

Dec 12, 2019 · Here, a detailed protocol for the detection and quantitative analysis of in vivo protein–protein proximity of Sox2 and Oct4 using the proximity-utilizing biotinylation (PUB) method is described.

The samples from experiments with the co-expression of BAP-Sox2 and BirA-Oct4 in HEK293T cells showed a high level of biotinylation (86 ± 6 and 71 ± 5 for different biotinylation times). This is due to the presence of DNA binding domains, HMG present in Sox2 and POU in Oct4, which recognize Utf1 or other motifs [ 36 ] and result in close ...

Jan 16, 2020 · Here, a detailed protocol for the detection and quantitative analysis of in vivo protein-protein proximity of Sox2 and Oct4 using the proximity-utilizing biotinylation (PUB) method is described.

The shotgun method was used to identify abundant proteins nonspecifically bound to the beads after washing. BAP-Sox2 and BAP-GFP fusion proteins were also identified, among other abundant proteins, and highlighted in yellow (Supplementary Tables S1–S4).

Feb 16, 2022 · Here we describe the rationale, design, and protocol for the new biotin acceptor peptides BAP1070 and BAP1108 using modular assembling of biotin acceptor fragments, DNA sequencing, transient expression of proteins in cells, and Western blotting methods.

Dec 30, 2022 · Treatment with an anti-SOX2 antibody on the third blot indicates recombinant BAP-SOX2 and the absence of a detectable amount of endogenous SOX2. BAP adds a mass shift of 3 kDa, and only a single band was observed on the anti-SOX2 blot in lane 2.