Journal: Ecotoxicology and environmental safety

Article Title: PRKAA1 induces aberrant mitophagy in a PINK1/Parkin-dependent manner, contributing to fluoride-induced developmental neurotoxicity.

doi: 10.1016/j.ecoenv.2023.114772

Figure Lengend Snippet: Fig. 5. NaF exposure induces elevated phos phorylation of PRKAA1 expression in vivo and in vitro. (A) The hierarchical cluster heatmap of phosphorylated proteins in rat hippocampus. (B) Statistics of different phosphorylation sites and proteins in rat hippocampus. (C) Volcano plot of differentially phosphorylated proteins in rat hippocampus. The criteria for significance were set at logarithmic fold change (log2 FC) of > 1.2 or < 0.8 (P < 0.05). (D) Phosphorylation of PRKAA1 protein in rat hippocampus. (E-F) Representative western blot and relative quan tifications of PRKAA1 and p-PRKAA1 in rat hippocampus. (G-H) Representative western blot and relative quantifications of PRKAA1 and p-PRKAA1 in SH-SY5Y cells. All experiments were performed independently and repeated 3 times. The data were presented as the means ± SD. *P < 0.05 compare with the control group.

Article Snippet: PRKAA1, p-PRKAA1 (Thr172), Parkin, and Cyt C antibodies were obtained from Boster Biotech (China).

Techniques: Expressing, In Vivo, In Vitro, Phospho-proteomics, Western Blot, Control

Journal: Ecotoxicology and environmental safety

Article Title: PRKAA1 induces aberrant mitophagy in a PINK1/Parkin-dependent manner, contributing to fluoride-induced developmental neurotoxicity.

doi: 10.1016/j.ecoenv.2023.114772

Figure Lengend Snippet: Fig. 6. DM inhibits aberrant mitophagy by pre venting the phosphorylation of PRKAA1 in vitro. (A-B) Representative western blot and relative quantifications of PRKAA1and p-PRKAA1 in SH- SY5Y cells. (C-D) Representative western blot and relative quantifications of PINK1 and Parkin in SH-SY5Y cells. (E-F) Representative images and relative quantifications of immunofluores cence staining of PINK1 and Mito-tracker, after DM intervention. The scale bar represents 50 µm. (G-H) Representative western blot and relative quantifications of TOMM-20 in SH-SY5Y cells. All experiments were performed independently and repeated 3 times. The data were presented as the means ± SD. *P < 0.05 compare with the control group, @P < 0.05 compare with the NaF group.

Article Snippet: PRKAA1, p-PRKAA1 (Thr172), Parkin, and Cyt C antibodies were obtained from Boster Biotech (China).

Techniques: Phospho-proteomics, In Vitro, Western Blot, Staining, Control

Journal: Frontiers in Oncology

Article Title: Angustoline Inhibited Esophageal Tumors Through Regulating LKB1/AMPK/ELAVL1/LPACT2 Pathway and Phospholipid Remodeling

doi: 10.3389/fonc.2020.01094

Figure Lengend Snippet: The levels of LKB1, AMPK, ELAVL1, LPCAT2, and β-actin proteins. (A) Expression of these proteins in 30 normal tissue samples and 30 esophageal cancer tissue samples. (B) Densitometric quantitations for normalized proteins relative to β-actin (%) in (A) .

Article Snippet: LKB1 siRNA (10 μM), AMPK activator (10 μM), LPCAT2 siRNA (10 μM), AMPK siRNA (10 μM) and angustoline (1 mg/L) were added into the wells, respectively. siRNA fragment treatment complex was prepared as follows, (1) Transfectant reagent (4 μL/well, Santa Cruz Biotechnology) was diluted in 1 mL of fresh 1,640 medium. (2) The siRNA fragment (4 μg/well) was diluted in 1 mL of fresh medium. (3) The compounds in (1) and (2) were mixed together and the DNA-liposome complex (2 mL per well) was added into the cells.

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: Angustoline Inhibited Esophageal Tumors Through Regulating LKB1/AMPK/ELAVL1/LPACT2 Pathway and Phospholipid Remodeling

doi: 10.3389/fonc.2020.01094

Figure Lengend Snippet: The levels of LKB1, AMPK, ELAVL1, LPCAT2, and β-actin proteins. (A) Expression of these proteins in KYSE-450 cells treated with AMPK activator or LPCAT2 siRNA fragment. (B) Expression of these proteins in KYSE-450 cells treated with AMPK siRNA or angustoline. (C) Expression of these proteins in KYSE-450 cells treated with LKB1 siRNA or angustoline. (D) Densitometric quantitations for normalized proteins relative to β-actin (%) in (A) . (E) Densitometric quantitations for normalized proteins relative to β-actin (%) in (B) . (F) Densitometric quantitations for normalized proteins relative to β-actin (%) in (C) . The data were represented as mean ± SD, * p < 0.05, compared with the control. n = 3.

Article Snippet: LKB1 siRNA (10 μM), AMPK activator (10 μM), LPCAT2 siRNA (10 μM), AMPK siRNA (10 μM) and angustoline (1 mg/L) were added into the wells, respectively. siRNA fragment treatment complex was prepared as follows, (1) Transfectant reagent (4 μL/well, Santa Cruz Biotechnology) was diluted in 1 mL of fresh 1,640 medium. (2) The siRNA fragment (4 μg/well) was diluted in 1 mL of fresh medium. (3) The compounds in (1) and (2) were mixed together and the DNA-liposome complex (2 mL per well) was added into the cells.

Techniques: Expressing, Control

Journal: Frontiers in Oncology

Article Title: Angustoline Inhibited Esophageal Tumors Through Regulating LKB1/AMPK/ELAVL1/LPACT2 Pathway and Phospholipid Remodeling

doi: 10.3389/fonc.2020.01094

Figure Lengend Snippet: In vivo effects of AMPK activator/AMPK antibody/Angustoline on KYSE450 tumor-bearing nude mice. (A) Treatment of AMPK antibody/AMPK activator/Angustoline/(AMPK antibody + Angustoline)/(AMPK activator + Angustoline) on the size of KYSE450 tumors. (B) Relative tumor volume, which was calculated by each tumor volume. * p < 0.05, comparing with the control, n = 5. (C) Tumor suppression rate, which was calculated by each tumor weight. * p < 0.05, comparing with the control, n = 5. (D) Relative tumor proliferation rate, which was calculated by relative tumor volumes of different groups. * p < 0.05, comparing with the control, n = 5.

Article Snippet: LKB1 siRNA (10 μM), AMPK activator (10 μM), LPCAT2 siRNA (10 μM), AMPK siRNA (10 μM) and angustoline (1 mg/L) were added into the wells, respectively. siRNA fragment treatment complex was prepared as follows, (1) Transfectant reagent (4 μL/well, Santa Cruz Biotechnology) was diluted in 1 mL of fresh 1,640 medium. (2) The siRNA fragment (4 μg/well) was diluted in 1 mL of fresh medium. (3) The compounds in (1) and (2) were mixed together and the DNA-liposome complex (2 mL per well) was added into the cells.

Techniques: In Vivo, Control