Journal: Molecular and cellular biochemistry

Article Title: Early-life bisphenol A exposure causes neuronal pyroptosis in juvenile and adult male rats through the NF-κB/IL-1β/NLRP3/caspase-1 signaling pathway: exploration of age and dose as effective covariates using an in vivo and in silico modeling approach.

doi: 10.1007/s11010-024-05039-4

Figure Lengend Snippet: Fig. 8 The molecular docking analysis of BPA and neuro- inflammatory, autophagic as well as, pyroptotic molecules displaying 2D and 3D binding interactions of BPA [PubChem CID: 6623] against A NF-kB [PDB ID: 1NFK], B IL-1β [PDB ID: 2MIB], C IL-2 [PDB ID: 4YQX], D IL-12 [PDB ID: 3HMX], E COX-2 [PDB ID: 1PXX], F NLRP3 [PDB ID: 7vtq], G Beclin-1 [PDB ID: 2PON], H LC3A [PDB ID: 6TBE], I LC3B [PDB ID: 5XAC], J Caspase-1 [PDB ID: 6VIE]. The green dotted lines denote hydrogen bonds between ligand and aminoacids, whereas bink/purple dotted lines repre- sent hydrophobic interactions. Electrostatic interactions are shown as orange dotted lines. The red dotted line indicate an unfavorable donor-donor

Article Snippet: Fine Test (cat no. ER1965) provided the NLRP3 kit, LSBio (cat no. LS-F9917; LS-F19802) provided the LC3A and LC3B kits, respectively, and Cusabio (cat no. CSB-EL002658RA provided the beclin-1 kit.

Techniques: Binding Assay

Journal: Genes & Development

Article Title: JNK regulates FoxO-dependent autophagy in neurons

doi: 10.1101/gad.1984311

Figure Lengend Snippet: JNK deficiency in neurons causes increased autophagy. (A) Wild-type (control) and JNKTKO CGNs infected with Ad-cre at 3 DIV were harvested at 10 DIV to prepare protein extracts that were examined using antibodies to LC3b, p62/SQSTM1, and α-Tubulin. (B) Extracts prepared from control and JNKTKO CGNs were examined by immunoblot analysis by probing with antibodies to Bcl-XL, Bnip3, Beclin-1, and α-Tubulin. Coimmunoprecipitation assays were performed by immunoblot analysis of Bcl-XL immunoprecipitates. (C) Extracts prepared from control and JNKTKO CGNs were examined by immunoblot (IB) analysis by probing with antibodies to AKT, pSer308-AKT, pSer473-AKT, FoxO1, pSer246-FoxO1, and α-Tubulin. CDK2 activity was measured in an immunecomplex kinase assay (KA) using Rb as the substrate. The relative CDK2 activity is indicated below. (D) Control and JNKTKO CGNs were stained with βIII-Tubulin and LC3b antibodies and examined by fluorescence microscopy. Bar, 10 μm. (E) Gene expression in CGNs was examined by quantitative RT–PCR analysis of mRNA and normalized to the amount of Gapdh mRNA in each sample (mean ± SD; n = 3). Statistically significant differences are indicated. (*) P < 0.05. (F) Control and JNKTKO CGNs were stained with DAPI and antibodies to FoxO1 and βIII-Tubulin. The neurons were examined by fluorescence microscopy. The merged image represents colocalization of FoxO1 with DAPI. Bar, 10 μm.

Article Snippet: TaqMan assays were used to quantitate Atg3 (Mm00471287_m1), Atg5 (Mm00504340_m1), Atg7 (Mm00512209_m1), Atg8/Lc3b (Mm00782868_m1), Atg12 (Mm00503201_m1), Beclin-1 (Mm01265461_m1), Bim (Mm01975020_s1), Bnip3 (Mm01275601_g1), FoxO1 (Mm00490672_M1), Gapdh (4352339E), Kif3a (Mm00492876_m1), Kif5a (Mm00515258_m1), Kif5b (Mm00515276_m1), and Kif5c (Mm00515265_m1) (Applied Biosystems).

Techniques: Control, Infection, Western Blot, Activity Assay, Kinase Assay, Staining, Fluorescence, Microscopy, Gene Expression, Quantitative RT-PCR

Journal: Genes & Development

Article Title: JNK regulates FoxO-dependent autophagy in neurons

doi: 10.1101/gad.1984311

Figure Lengend Snippet: Effect of RNAi-mediated knockdown of Beclin-1 on autophagy and survival of JNKTKO neurons. (A) Wild-type (control) and Jnk1LoxP/LoxP Jnk2−/− Jnk3−/− (JNKTKO) neurons infected with Ad-cre at 3 DIV were transfected at 7 DIV with Beclin-1 siRNA or control siRNA. The expression of Beclin-1 mRNA was examined at 11 DIV by quantitative RT–PCR analysis of mRNA and normalized to the amount of Gapdh mRNA in each sample (mean ± SD; n = 3). Statistically significant differences are indicated. (*) P < 0.05. (B) Control and JNKTKO neurons transfected with scrambled sequence or Beclin-1 siRNA were examined at 11 DIV by immunoblot analysis with antibodies to LC3b, p62/SQSTM1, and α-Tubulin. (C) The survival of RNAi transfected control and JNKTKO neurons at 11 DIV was quantitated (mean ± SD; n = 20). Statistically significant differences are indicated. (*) P < 0.05.

Article Snippet: TaqMan assays were used to quantitate Atg3 (Mm00471287_m1), Atg5 (Mm00504340_m1), Atg7 (Mm00512209_m1), Atg8/Lc3b (Mm00782868_m1), Atg12 (Mm00503201_m1), Beclin-1 (Mm01265461_m1), Bim (Mm01975020_s1), Bnip3 (Mm01275601_g1), FoxO1 (Mm00490672_M1), Gapdh (4352339E), Kif3a (Mm00492876_m1), Kif5a (Mm00515258_m1), Kif5b (Mm00515276_m1), and Kif5c (Mm00515265_m1) (Applied Biosystems).

Techniques: Knockdown, Control, Infection, Transfection, Expressing, Quantitative RT-PCR, Sequencing, Western Blot