mouse monoclonal anti sqstm1  (Novus Biologicals)

Journal: Bioactive Materials

Article Title: A foam cell-targeted lipophagy restoration strategy stabilizes vulnerable atherosclerotic plaques

doi: 10.1016/j.bioactmat.2026.02.041

Figure Lengend Snippet: In vitro evaluation of foam cell lipid accumulation and lipophagy activation following OPN-HMCN@MLT treatment. ( A - C ) ORO and BODIPY staining images and corresponding quantification of ORO and BODIPY positive areas of RAW264.7 cells under different stimulations (n = 5, scale bar for ORO: 100 μm, scale bar for BODIPY: 20 μm). ( D ) Bio-TEM images of RAW264.7 cells post various treatments (n = 5, scale bars 1.0 μm). Green arrows indicate nanoparticles. ( E , F ) Morphometric analysis determined the mean number and area (μm 2 ) of LDs per cell section. ( G ) Confocal images depicting lipophagy flux in foam cells following different treatments (n = 5 biological replicates, scale bars: 10 μm). ( H - J ) The quantities of acidified autophagosomes (GFP-RFP+), neutral autophagosomes (GFP + RFP+), and LDs labeled with BODIPY were measured per cell for each condition. (K to N) Representative Western blot images and quantitative analysis of LC3, LAMP1, and P62 expression in foam cells. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001.

Article Snippet: To block nonspecific binding, membranes were incubated with 5% skim milk for 1 h. Thereafter, membranes were incubated overnight at 4 °C with primary antibodies against ABCA1, ABCG1, ACOX1, CPT1A, LC3 (ab192890, 1:2000, abcam), LAMP1 (84658-5-RR, 1:8000, Proteintech), PPARα (66826-1-Ig, 1:3000, Proteintech), PPARγ (66936-1-Ig, 1:10000, Proteintech), P62 (18420-1-AP, 1:10000, Proteintech), MCAD (55210-1-AP, 1:3000, Proteintech), LCAD (17526-1-AP, 1:10000, Proteintech), tubulin (80762-1-RR, 1:10000, Proteintech), GAPDH (60004-1-Ig, 1:50000, Proteintech), and β-actin (66009-1-Ig, 1:20000, Proteintech).

Techniques: In Vitro, Activation Assay, Staining, Labeling, Western Blot, Expressing

Journal: Oncology Reports

Article Title: Platycodin D sensitizes head and neck squamous cell carcinoma to cisplatin by inducing autophagy arrest

doi: 10.3892/or.2026.9088

Figure Lengend Snippet: PD and cisplatin combination treatment increases autophagy marker levels in head and neck squamous cell carcinoma cells. (A) HSC3 and (C) FaDu cells were treated with PD (10 µM), cisplatin (10 µM) or combination treatment for 48 h. Cells were stained with LC3B antibody and analyzed using confocal microscopy. (B) In HSC3 cells, p62 immunofluorescence was increased following combination treatment compared with single treatments. (D) In FaDu cells, p62 staining similarly showed higher p62 levels in the combination treatment group than in the single treatment groups. Nuclei were stained with DAPI (blue), LC3B was stained red and p62 was stained green. Fluorescence intensity and puncta quantification were analyzed using ZEN software (Zeiss AG). Scale bar, 50 µm. Data are presented as the mean ± SD (n=3). *P<0.05, **P<0.01, ***P<0.001. PD, Platycodin D.

Article Snippet: The following antibodies were used: Anti-LC3A/B (1:1,000; cat. no. 12741S; Cell Signaling Technology, Inc.), anti-SQSTM1/p62 (1:1,000; cat. no. 5114S; Cell Signaling Technology, Inc.), anti-poly(ADP-ribose) polymerase (PARP; 1:1,000; cat. no. 9542S; Cell Signaling Technology, Inc.), anti-caspase 3 (1:1,000; cat. no. 9662S; Cell Signaling Technology, Inc.) and anti-β-actin (1:5,000; cat. no. 47778; Santa Cruz Biotechnology, Inc.).

Techniques: Marker, Staining, Confocal Microscopy, Immunofluorescence, Fluorescence, Software

Journal: Oncology Reports

Article Title: Platycodin D sensitizes head and neck squamous cell carcinoma to cisplatin by inducing autophagy arrest

doi: 10.3892/or.2026.9088

Figure Lengend Snippet: PD and cisplatin combination treatment induces autophagy arrest by inhibiting autophagosome degradation. HSC3 and FaDu cells were treated with PD (10 µM), cisplatin (10 µM) or combination treatment for 48 h, with or without BafA1. (A) Western blotting was performed to evaluate the expression levels of LC3A/B and p62. β-actin was used as a loading control. (B) Densitometric analysis of LC3A/B and p62 protein levels normalized to β-actin, performed using ImageJ software. Statistical significance was determined by one-way ANOVA followed by Tukey's post hoc test. Data are presented as the mean ± SD (n=3). *P<0.05, **P<0.01. BafA1, Bafilomycin A1; PD, Platycodin D.

Article Snippet: The following antibodies were used: Anti-LC3A/B (1:1,000; cat. no. 12741S; Cell Signaling Technology, Inc.), anti-SQSTM1/p62 (1:1,000; cat. no. 5114S; Cell Signaling Technology, Inc.), anti-poly(ADP-ribose) polymerase (PARP; 1:1,000; cat. no. 9542S; Cell Signaling Technology, Inc.), anti-caspase 3 (1:1,000; cat. no. 9662S; Cell Signaling Technology, Inc.) and anti-β-actin (1:5,000; cat. no. 47778; Santa Cruz Biotechnology, Inc.).

Techniques: Western Blot, Expressing, Control, Software

Journal: Oncology Reports

Article Title: Platycodin D sensitizes head and neck squamous cell carcinoma to cisplatin by inducing autophagy arrest

doi: 10.3892/or.2026.9088

Figure Lengend Snippet: PD and cisplatin combination treatment increases autophagy marker levels in head and neck squamous cell carcinoma cells. (A) HSC3 and (C) FaDu cells were treated with PD (10 µM), cisplatin (10 µM) or combination treatment for 48 h. Cells were stained with LC3B antibody and analyzed using confocal microscopy. (B) In HSC3 cells, p62 immunofluorescence was increased following combination treatment compared with single treatments. (D) In FaDu cells, p62 staining similarly showed higher p62 levels in the combination treatment group than in the single treatment groups. Nuclei were stained with DAPI (blue), LC3B was stained red and p62 was stained green. Fluorescence intensity and puncta quantification were analyzed using ZEN software (Zeiss AG). Scale bar, 50 µm. Data are presented as the mean ± SD (n=3). *P<0.05, **P<0.01, ***P<0.001. PD, Platycodin D.

Article Snippet: The cells were incubated overnight at 4°C with primary antibodies against LC3B (1:1,000; cat. no. 2775S; Cell Signaling Technology, Inc.) and sequestosome 1 (SQSTM1)/p62 (1:1,000; cat. no. 5114S; Cell Signaling Technology, Inc.).

Techniques: Marker, Staining, Confocal Microscopy, Immunofluorescence, Fluorescence, Software

Journal: Oncology Reports

Article Title: Platycodin D sensitizes head and neck squamous cell carcinoma to cisplatin by inducing autophagy arrest

doi: 10.3892/or.2026.9088

Figure Lengend Snippet: PD and cisplatin combination treatment induces autophagy arrest by inhibiting autophagosome degradation. HSC3 and FaDu cells were treated with PD (10 µM), cisplatin (10 µM) or combination treatment for 48 h, with or without BafA1. (A) Western blotting was performed to evaluate the expression levels of LC3A/B and p62. β-actin was used as a loading control. (B) Densitometric analysis of LC3A/B and p62 protein levels normalized to β-actin, performed using ImageJ software. Statistical significance was determined by one-way ANOVA followed by Tukey's post hoc test. Data are presented as the mean ± SD (n=3). *P<0.05, **P<0.01. BafA1, Bafilomycin A1; PD, Platycodin D.

Article Snippet: The cells were incubated overnight at 4°C with primary antibodies against LC3B (1:1,000; cat. no. 2775S; Cell Signaling Technology, Inc.) and sequestosome 1 (SQSTM1)/p62 (1:1,000; cat. no. 5114S; Cell Signaling Technology, Inc.).

Techniques: Western Blot, Expressing, Control, Software