p62  (Bioss)

Journal: Frontiers in Endocrinology

Article Title: Spatio-temporal dynamics of autophagy-associated genes in macrophage-driven atherosclerosis: an integrated omics and experimental study

doi: 10.3389/fendo.2026.1764263

Figure Lengend Snippet: Expression detection of key genes in AS macrophages. (A) statistical graph of SNX5, SMG1, GSK3A mRNA expression level. (B) The SNX5, SMG1, GSK3A protein expression levels: Left, typical western blots, Right, statistical graph. (C) Protein expression of autophagy markers LC3 and p62. * P < 0.05, ** P < 0.01, *** P < 0.001 vs. Raw264.7.

Article Snippet: In addition, the protein expression levels of LC3 (Affinity, AF7001) and P62 (BIOSS, bs-8878R) were assessed to evaluate autophagic flux, with β-actin (Proteintech, 66009-1-Ig) serving as the loading control for normalization.

Techniques: Expressing, Western Blot

Journal: bioRxiv

Article Title: Dysregulation of a novel autophagosome-mitochondria contact contributes to autophagy dysfunction and neurodegeneration in tauopathy

doi: 10.64898/2026.03.23.713823

Figure Lengend Snippet: Autophagosome/autophagic vacuole (AV)-mitochondria (Mito) contact hyper-tethering and AV retrograde transport defects in tauopathy axons. ( A and B ) Representative transmission electron microscopy (TEM) images ( A ) and quantitative analysis ( B ) of 8-month-old tauP301S Tg (PS19) hippocampi. The number of presynaptic AVs, the percentage of terminals containing AVs, and the number of AVs or mitochondria in AV-Mito contacts were quantified and normalized to or compared to those in non-Tg littermate controls. Data were quantified from a total number of presynaptic terminals ( n ) as indicated in parentheses ( B ) from three mice per group. Arrows: AV-Mito contacts; AV: autophagic vacuole; M: mitochondrion. ( C - F ) Representative time-lapse images and kymographs ( C ) and quantitative analysis ( D - F ) of non-Tg and tauP301S Tg axons after 24-hour trehalose (100 mM) incubation. AV-Mito contact duration and its frequency, the contact number per 100 μm axonal length, AV percentage in contacts, and the relative motility of AVs were quantified and compared between the two groups, respectively. Data were collected from three dissections: the total numbers of neurons ( n ) and AV-Mito contacts ( c ) are indicated in parentheses ( D - F ). Arrows: AVs (time-lapse images in top C ); AV-Mito contacts (kymographs in bottom C ). ( G ) Correlative light and electron microscope (CLEM) and cryo-electron tomography (cryo-ET) analysis of AV-Mito contacts in tauopathy axons. Low-magnification cryo-EM image of a tauP301S Tg neuron showing a region of the axonal process ( G1 ). CLEM of the region outlined by the white box in G1 , showing fluorescence images of mRFP-LC3-labeled AVs ( G2 ) and MitoView Green-marked mitochondria ( G3 ). The cryo-EM image of the same region acquired at 3,600 × magnification ( G4 ) is overlaid with the fluorescence image ( G5 ). The white dashed box indicates the potential AV-Mito contact site selected for tilt-series acquisition. Slice view of the reconstructed tomogram ( G6 ) with membrane annotations of a mitochondrion (green), an AV (red), synaptic vesicles (purple), and membranes (light grey) ( G7 ). Zoomed-in view of the AV-Mito contact site showed densities bridging the two membranes ( G8 , white arrows). Data were expressed as the mean ± SEM and analyzed using linear mixed-effects models. Scale bars: 100 nm ( A ) and 10 μm ( C ).

Article Snippet: Sources of antibodies or reagents are as follows: polyclonal antibodies against LC3 (Cell Signaling Technology, Cat# 2775), AMPKα (Cell Signaling Technology, Cat# 2532), p62/SQSTM1 (Abnova, Cat# H00008878-M01), NeuN (Millipore/Sigma, Cat# ABN78), TBC1D15 (Abcam, Cat# ab121396), Tau5 (DAKO, Cat# A0024), and synaptophysin/SYP (Abcam, Cat# ab32127); monoclonal antibodies against phospho-AMPKα (Cell Signaling Technology, Cat# 2535), AT8 (ThermoFisher Scientific, Cat# MN1020), syntaxin 1/STX1 (Santa Cruz Biotechnology, Cat# sc-12736), Rab7 (Sigma, Cat# R8779), GAPDH (Sigma, Cat# CB1001), TOM20 (Abcam, Cat# ab186734), mCherry (Takara, Cat# 632543), and Alexa fluor 488- (Cat# A-11017; Cat# A-11070), 546- (Cat# A-11018; Cat# A-11071), and 647- (Cat# A-21237; Cat# A-21246) conjugated secondary antibodies (Invitrogen); 10-NCP (VWR, Cat# 80017-188), trehalose (Cat# T0167), cycloheximide (Cat# 01810), rotenone (Cat# 557368), antimycin A (Cat# 1397-94-0), AICAR (Cat# A9978), CC (Cat# 171264), epoxomicin (Cat# 324800), CID 1067700 (Cat# SML0545), DMSO (Cat# D2650), and Sarkosyl (Cat# L9150) (Sigma); TBC1D15 shRNA (Cat# sc-154093-SH), Fis1 shRNA (Cat# sc-60644-SH), and control shRNA (Cat# sc-108060) (Santa Cruz Biotechnology); MitoView TM Green (Cat# 70054) (biotium); LentiBrite RFP-LC3 Lentiviral Biosensor (Cat# 17-10143) (Sigma).

Techniques: Transmission Assay, Electron Microscopy, Incubation, Microscopy, Tomography, Cryo-EM Sample Prep, Fluorescence, Labeling, Membrane

Journal: Frontiers in Microbiology

Article Title: ULK1 mediated autophagy in airway cells during Aspergillus infection

doi: 10.3389/fmicb.2026.1756294

Figure Lengend Snippet: Increased LC3-II expression and monolayer membrane-coated autophagosome formation in Beas2B cells following conidia infection. (A) Expression of LC3-I and LC3-II in Beas2B following infected by B5233 wild-type live conidia at 0, 2, 4, 6, 8, and 10 h. (B) Relative expression levels of LC3-I and LC3-II at the various time points. (C) Fluorescence microscopy images of GFP-LC3 (green), mCherry-labeled conidia (red), and DAPI staining (blue) in Beas2B cells following infected by conidia at 0, 4, 6, and 8 h. (D) Quantification of cells with LC3 + vesicles and average number of LC3 + vesicles per cell at 4, 6, and 8 h. (E) Electron microscopy images of cellular structural changes over time at 2, 4, 8, and 12 h. The magnification progressively increases from left to right, with each subsequent column of images depicting the area enclosed within the red box of the preceding image. The red arrow in each image points to the monolayer membrane encapsulating the A. fumigatus spores. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: LC3 Rabbit Polyclonal antibody(#14600-1-AP, 1:1000), Rubicon Rabbit Polyclonal Antibody (#21444-1-AP, 1:1000), GAPDH Polyclonal antibody(#10494-1-AP, 1:2000) were purchased from proteintech, Phospho-UKL1(Ser555) Rabbit monoclonal antibody (#5869, 1:500), Phospho-UKL1(Ser757) Rabbit monoclonal antibody (#6888, 1:500), ULK1 rabbit monoclonal antibody (#8054, 1:1000), SQSTM1/p62 antibody (#5114, 1:1000), Atg5 Rabbit monoclonal antibody (#12994, 1:1000), Dectin-1 Rabbit monoclonal antibody (#60128, 1:1000), AMPKalpha Antibody (#2532, 1:1000), Phospho-AMPKalpha (Thr172) Rabbit monoclonal antibody(#50081, 1:1000), β -tubulin Rabbit monoclonal antibody(#2128, 1:2000) were purchased from Cell Signaling Technology (USA).

Techniques: Expressing, Membrane, Infection, Fluorescence, Microscopy, Labeling, Staining, Electron Microscopy

Journal: Frontiers in Microbiology

Article Title: ULK1 mediated autophagy in airway cells during Aspergillus infection

doi: 10.3389/fmicb.2026.1756294

Figure Lengend Snippet: ULK1 and CR3 receptor modulated conidia-induced autophagy in Beas2B cells. (A) Expression levels of ULK1, p-ULK1 (Ser555), and p-ULK1 (Ser757) in Beas2B after internalization by B5233 wild-type live conidia at 0, 1, 2, 4, 6, and 8 h. (B) Expression levels of LC3-I and LC3-II in ULK1-silenced Beas2B cells after internalization by conidia. (C) Expression levels of LC3-I, LC3-II, ULK1, p-ULK1 (Ser555), and p-ULK1 (Ser757) in Beas2B cells treated with 2 μM and 5 μM SBI-0206965 inhibitor targeting ULK1/2 with conidia infection. (D) Expression levels of LC3-I and LC3-II in Beas2B cells with CR3 knockdown or overexpression after internalization by conidia. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: LC3 Rabbit Polyclonal antibody(#14600-1-AP, 1:1000), Rubicon Rabbit Polyclonal Antibody (#21444-1-AP, 1:1000), GAPDH Polyclonal antibody(#10494-1-AP, 1:2000) were purchased from proteintech, Phospho-UKL1(Ser555) Rabbit monoclonal antibody (#5869, 1:500), Phospho-UKL1(Ser757) Rabbit monoclonal antibody (#6888, 1:500), ULK1 rabbit monoclonal antibody (#8054, 1:1000), SQSTM1/p62 antibody (#5114, 1:1000), Atg5 Rabbit monoclonal antibody (#12994, 1:1000), Dectin-1 Rabbit monoclonal antibody (#60128, 1:1000), AMPKalpha Antibody (#2532, 1:1000), Phospho-AMPKalpha (Thr172) Rabbit monoclonal antibody(#50081, 1:1000), β -tubulin Rabbit monoclonal antibody(#2128, 1:2000) were purchased from Cell Signaling Technology (USA).

Techniques: Expressing, Infection, Knockdown, Over Expression

Journal: Frontiers in Microbiology

Article Title: ULK1 mediated autophagy in airway cells during Aspergillus infection

doi: 10.3389/fmicb.2026.1756294

Figure Lengend Snippet: Model of a novel ULK1-mediated LAP-like pathway in airway cells during A. fumigatus conidial internalization. During the interaction between A. fumigatus conidia and airway cells, several changes occur: intracellular LC3-II levels, phosphorylation of AMPK and ULK1, and reactive oxygen species (ROS) content all increase. Additionally, the release of cytokines—including IL-6, IL-8, and MCP-1—also rises. Inhibiting ULK1 activity or silencing its expression can suppress the increase in LC3-II levels and cytokine release triggered by A. fumigatus . Notably, common fungal polysaccharides and Dectin-1 did not impact this process, but the loss of complement receptor 3 elevated both basal and conidia-induced autophagy, correlating with increased AMPK expression. Dashed lines indicate signal pathway interactions hypothesized to occur in airway cells.

Article Snippet: LC3 Rabbit Polyclonal antibody(#14600-1-AP, 1:1000), Rubicon Rabbit Polyclonal Antibody (#21444-1-AP, 1:1000), GAPDH Polyclonal antibody(#10494-1-AP, 1:2000) were purchased from proteintech, Phospho-UKL1(Ser555) Rabbit monoclonal antibody (#5869, 1:500), Phospho-UKL1(Ser757) Rabbit monoclonal antibody (#6888, 1:500), ULK1 rabbit monoclonal antibody (#8054, 1:1000), SQSTM1/p62 antibody (#5114, 1:1000), Atg5 Rabbit monoclonal antibody (#12994, 1:1000), Dectin-1 Rabbit monoclonal antibody (#60128, 1:1000), AMPKalpha Antibody (#2532, 1:1000), Phospho-AMPKalpha (Thr172) Rabbit monoclonal antibody(#50081, 1:1000), β -tubulin Rabbit monoclonal antibody(#2128, 1:2000) were purchased from Cell Signaling Technology (USA).

Techniques: Phospho-proteomics, Activity Assay, Expressing

Journal: Frontiers in Oncology

Article Title: Study on the mechanism of 18β-glycyrrhetinic acid inhibiting the proliferation of renal cancer cells by inducing autophagy through the miR-27a-5p/LC3 axis

doi: 10.3389/fonc.2026.1762770

Figure Lengend Snippet: The effect of miR-27a-5p on the proliferation phenotype of human renal cancer cells. (A) The effect of miR-27a-5p on the clone formation ability of human renal cancer cells. (B) The effect of miR-27a-5p on the apoptosis of human renal cancer cells. (C) The effect of miR-27a-5p on human renal cancer cell cycle. (D) The effect of miR-27a-5p on the expression of LC3 protein in human renal cancer cells *p < 0.05, **p < 0.01, ***p < 0.001 indicate that the results are statistically significant.

Article Snippet: LC3 rabbit polyclonal antibody , Wuhan Sanying Biotechnology Co.Ltd. , Cat No. 14600-1-AP.

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: Study on the mechanism of 18β-glycyrrhetinic acid inhibiting the proliferation of renal cancer cells by inducing autophagy through the miR-27a-5p/LC3 axis

doi: 10.3389/fonc.2026.1762770

Figure Lengend Snippet: 18-GA regulates the effect of miR-27a-5p on the proliferation phenotype of human renal cancer cells. (A) 18-GA regulates the effect of miR-27a-5p on the clone formation ability of human renal cancer cells. (B) 18-GA regulates the effect of miR-27a-5p on the apoptosis of human renal cancer cells. (C) 18-GA regulates the effect of miR-27a-5p on the cell cycle of human renal cancer cells. (D) 18-GA regulates the effect of miR-27a-5p on the expression of LC3 protein in human renal cancer cells. *p < 0.05, **p < 0.01, ***p < 0.001, *p < 0.05, **p < 0.01, ***p < 0.001 indicate that the results are statistically significant.

Article Snippet: LC3 rabbit polyclonal antibody , Wuhan Sanying Biotechnology Co.Ltd. , Cat No. 14600-1-AP.

Techniques: Expressing