Journal: American journal of physiology. Gastrointestinal and liver physiology

Article Title: Pioglitazone Reduces Hepatic Alpha-1 Antitrypsin Accumulation Through Autophagy and AMPK Activation in Alpha-1 Antitrypsin Deficient Mice

doi: 10.1152/ajpgi.00272.2025

Figure Lengend Snippet: (A) Western blot analysis of autophagy-related proteins in HuhZ cells treated with pioglitazone (10 μM, 48 h), with or without Bafilomycin A1 (BafA1, 100 μM, 6 h). Protein levels of ATG5, p62, and LC3B were assessed. GAPDH serves as a loading control. (B) Quantification of p62 and LC3B-II band intensities from (A), normalized to GAPDH. Bars represent mean ± SD from three independent experiments. (C) Immunofluorescence staining of LC3B (green) in HuhZ cells treated with pioglitazone ± BafA1. Nuclei were counterstained with DAPI (blue). Insets show enlarged views of LC3B-positive puncta. (D) Western blot analysis of AMPK-mTOR signaling components in control and pioglitazone-treated HuhZ cells. Phosphorylation of AMPK, ULK1, and mTOR was assessed. (E) Quantification of p-AMPK, p-ULK1, and p-mTOR band intensities from (D), normalized to GAPDH. Bars show mean ± SD. (F) Immunofluorescence staining of p-AMPK (green) and actin (red) in control and pioglitazone-treated HuhZ cells. Nuclei were counterstained with DAPI (blue). Merged images show increased p-AMPK activation upon pioglitazone treatment.

Article Snippet: The blots were blocked in 5% non-fat dried milk and incubated overnight at 4°C with rabbit polyclonal antibodies against ATG5, AMPK, p-AMPK, ULK, p-ULK, mTOR, p-mTOR, PTEN, and p-PTEN (Cell Signaling Technology, Danvers, MA, USA); LC3 (Proteintech, Rosemont, IL); and AAT (Dako, Carpenteria, CA).

Techniques: Western Blot, Control, Immunofluorescence, Staining, Phospho-proteomics, Activation Assay

Journal: American journal of physiology. Gastrointestinal and liver physiology

Article Title: Pioglitazone Reduces Hepatic Alpha-1 Antitrypsin Accumulation Through Autophagy and AMPK Activation in Alpha-1 Antitrypsin Deficient Mice

doi: 10.1152/ajpgi.00272.2025

Figure Lengend Snippet: (A) Western blot analysis of liver lysates from control- and pioglitazone-treated Pi*Z mice (30 mg/kg/day, 12 weeks) showing levels of total AAT, ATG5, LC3B-I/II, and p62. GAPDH is shown as a loading control. (B) Quantification of Western blot band intensities from (A), normalized to GAPDH. Data represents SD from 4 mice per group. Statistical significance assessed by unpaired t-test. (C) Immunohistochemistry for LC3B in liver sections from control and pioglitazone-treated mice. The right panel shows higher magnification with arrows indicating LC3B-positive vacuoles. (D) Immunohistochemistry for p62 in liver sections from control and pioglitazone-treated mice. p62-positive aggregates are reduced in pioglitazone-treated livers. Scale bars: 100 μm (all panels).

Article Snippet: The blots were blocked in 5% non-fat dried milk and incubated overnight at 4°C with rabbit polyclonal antibodies against ATG5, AMPK, p-AMPK, ULK, p-ULK, mTOR, p-mTOR, PTEN, and p-PTEN (Cell Signaling Technology, Danvers, MA, USA); LC3 (Proteintech, Rosemont, IL); and AAT (Dako, Carpenteria, CA).

Techniques: Western Blot, Control, Immunohistochemistry