Journal: Frontiers in Pharmacology

Article Title: Berberine Protects Against NLRP3 Inflammasome via Ameliorating Autophagic Impairment in MPTP-Induced Parkinson’s Disease Model

doi: 10.3389/fphar.2020.618787

Figure Lengend Snippet: BBR mitigates autophagic impairment in MPTP-induced mice. (A) The representative immunohistochemical staining for MAP1LC3B in SN. (B) The number of MAP1LC3B positive cells in SN. Representative western blot bands (C) and the statistical graph (D) of MAP1LC3B and BECN1 in SN. (E) Transmission electron microscopy shown with autophagosomes (yellow arrows) in the SN. Data were expressed as the mean ± SD ( n = 6). * p < 0.05 compared with control group, # p < 0.05 compared with MPTP group, + p < 0.05 compared with MPTP + BBR group. MAP1LC3B, microtubule associated protein 1 light chain 3 beta; SN, substantia nigra; BECN1, beclin 1.

Article Snippet: The primary antibodies were incubated overnight at 4 °C included anti-TH (1:500; Santa Cruz, sc-25269), anti-solute carrier family 6 member 3 (SLC6A3) (1:1,000; ABclonal, A152360), anti-dopamine receptor D2 (DRD2) (1:1,000; ABclonal, A12930), anti-AIF1 (1:1,000; Santa Cruz, sc-32725), anti-GFAP (1:1,000; ABclonal, A14673), anti-NLRP3 (1:200; AdipoGen, AG-20B-0014-C100), anti-PYCARD (1:1,000; Immunoway, T0365), anti-CASP1 (1:1,000; ABclonal, A0964), anti-IL1B (1:1,000; ABclonal, A12688), anti-MAP1LC3B (1:1,000; ABclonal, A11282), anti-BECN1 (1:1,000; Cell Signaling Technology, 3738S) and anti-ACTB (1:3,000; ABclonal, AC026).

Techniques: Immunohistochemical staining, Staining, Western Blot, Transmission Assay, Electron Microscopy

Journal: Frontiers in Pharmacology

Article Title: Berberine Protects Against NLRP3 Inflammasome via Ameliorating Autophagic Impairment in MPTP-Induced Parkinson’s Disease Model

doi: 10.3389/fphar.2020.618787

Figure Lengend Snippet: BBR enhances autophagic activity in MPP + -treated BV2 cells. The representative immunofluorescent staining (A) and puncta (B) of MAP1LC3B in BV2 cells treated with MPP + at 200 μM and BBR at the concentration of 12.5, 25, and 50 μM (20 cells were analyzed per group for MAP1LC3B puncta counting). Representative western blots (C) and the statistical graph (D) of MAP1LC3B and BECN1 in BV2 cells treated with MPP + at 200 μM and BBR at the concentration of 12.5, 25, and 50 μM. The representative double-immunofluorescent staining (E) and puncta (E) of MAP1LC3B in BV2 cells treated with MPP + at 200 μM, BBR at 25 μM and 3-MA at 10 mM (20 cells were analyzed per group for MAP1LC3B puncta counting). The representative monodansylcadaverine staining (G) and statistical graph (H) of autophagic vesicles in BV2 cells treated with MPP + at 200 μM, BBR at 25 μM and 3-MA at 10 mM. Representative western blot bands (I) and the statistical graph (J) of MAP1LC3B and BECN1 in BV2 cells treated with MPP + at 200 μM, BBR at 25 μM and 3-MA at 10 mM. Data were expressed as the mean ± SD ( n = 3). * p < 0.05 compared with untreated group, # p < 0.05 compared with MPP + group, + p < 0.05 compared with MPP + + BBR group. MAP1LC3B, microtubule associated protein 1 light chain 3 beta; BECN1, beclin 1.

Article Snippet: The primary antibodies were incubated overnight at 4 °C included anti-TH (1:500; Santa Cruz, sc-25269), anti-solute carrier family 6 member 3 (SLC6A3) (1:1,000; ABclonal, A152360), anti-dopamine receptor D2 (DRD2) (1:1,000; ABclonal, A12930), anti-AIF1 (1:1,000; Santa Cruz, sc-32725), anti-GFAP (1:1,000; ABclonal, A14673), anti-NLRP3 (1:200; AdipoGen, AG-20B-0014-C100), anti-PYCARD (1:1,000; Immunoway, T0365), anti-CASP1 (1:1,000; ABclonal, A0964), anti-IL1B (1:1,000; ABclonal, A12688), anti-MAP1LC3B (1:1,000; ABclonal, A11282), anti-BECN1 (1:1,000; Cell Signaling Technology, 3738S) and anti-ACTB (1:3,000; ABclonal, AC026).

Techniques: Activity Assay, Staining, Concentration Assay, Western Blot

Journal: Molecular Therapy. Nucleic Acids

Article Title: A Discovery of Clinically Approved Formula FBRP for Repositioning to Treat HCC by Inhibiting PI3K/AKT/NF-κB Activation

doi: 10.1016/j.omtn.2019.12.023

Figure Lengend Snippet: Effects of FBRP on Expression Levels of Candidate Targets Involved in PI3K/AKT/NF-κB Signaling in Liver Tissues of DEN-Induced HCC Rats (A) Western blots of the candidate targets of FBRP against HCC involved in PI3K/AKT/NF-κB signaling. (B–M) Relative protein expression levels of (B) p-PI3K, (C) p-AKT, (D) IKκB, (E) p-IKκB, (F) the ratio of p-IKκB to IKκB, (G) NF-κB (p65), (H) p-NF-κB (p65), (I) the ratio of NF-κB (p65) to p-NF-κB (p65), (J) CCND1, (K) CCNE1, (L) CDK2, and (M) CDK4 in different groups are shown. Data are expressed as the mean ± SD. # p < 0.05, ## p < 0.01, ### p < 0.001, comparison with the normal group; @ p < 0.05, @@ p < 0.01, @@@ p < 0.001, comparison with the DEN-12w group; *p < 0.05, **p < 0.01, ***p < 0.001, comparison with the DEN-15w group; $ p < 0.05, $$ p < 0.01, $$$ p < 0.001, comparison with the DEN-18w group.

Article Snippet: The detailed information on these antibodies is as follows: p-PI3K (catalog no. 4249, Cell Signaling Technology, MA, USA), p-AKT (catalog no. 4060, Cell Signaling Technology, MA, USA), IKκB (catalog no. 8943, Cell Signaling Technology, MA, USA), p-IKκB (catalog no. YP0637, ImmunoWay, TX, USA), NF-κB (p65) (catalog no. YP0191, ImmunoWay, TX, USA), p-NF-κB (p65) (catalog no. 3033, Cell Signaling Technology, MA, USA), CCND1 (catalog no. 2978, Cell Signaling Technology, MA, USA), CCNE1 (catalog no. 20808, Cell Signaling Technology, MA, USA), CDK2 (catalog no. 10122-1-AP, Proteintech, IL, USA), CDK4 (catalog no. 11026-1-AP, Proteintech, IL, USA), and GAPDH (catalog no. 2118, Cell Signaling Technology, MA, USA).

Techniques: Expressing, Western Blot

Journal: Molecular Therapy. Nucleic Acids

Article Title: A Discovery of Clinically Approved Formula FBRP for Repositioning to Treat HCC by Inhibiting PI3K/AKT/NF-κB Activation

doi: 10.1016/j.omtn.2019.12.023

Figure Lengend Snippet: Effects of FBRP on Expression Levels of Candidate Targets Involved in PI3K/AKT/NF-κB Signaling in the HCC Cell Line Huh7 (A) Western blots of the candidate targets of FBRP against HCC involved in PI3K/AKT/NF-κB signaling in different groups. (B–M) Relative protein expression levels of (B) p-PI3K, (C) p-AKT, (D) IKκB, (E) p-IKκB, (F) the ratio of p-IKκB to IKκB, (G) NF-κB (p65), (H) p-NF-κB (p65), (I) the ratio of NF-κB (p65) to p-NF-κB (p65), (J) CCND1, (K) CCNE1, (L) CDK2, and (M) CDK4 in different groups are shown. Data are expressed as the mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, comparison with the Huh7 group; # p < 0.05, ## p < 0.01, ### p < 0.001, comparison with the Huh7-FBRP treatment group; @ p < 0.05, @@ p < 0.01, @@@ p < 0.001, comparison with the Huh7-LY294002 group.

Article Snippet: The detailed information on these antibodies is as follows: p-PI3K (catalog no. 4249, Cell Signaling Technology, MA, USA), p-AKT (catalog no. 4060, Cell Signaling Technology, MA, USA), IKκB (catalog no. 8943, Cell Signaling Technology, MA, USA), p-IKκB (catalog no. YP0637, ImmunoWay, TX, USA), NF-κB (p65) (catalog no. YP0191, ImmunoWay, TX, USA), p-NF-κB (p65) (catalog no. 3033, Cell Signaling Technology, MA, USA), CCND1 (catalog no. 2978, Cell Signaling Technology, MA, USA), CCNE1 (catalog no. 20808, Cell Signaling Technology, MA, USA), CDK2 (catalog no. 10122-1-AP, Proteintech, IL, USA), CDK4 (catalog no. 11026-1-AP, Proteintech, IL, USA), and GAPDH (catalog no. 2118, Cell Signaling Technology, MA, USA).

Techniques: Expressing, Western Blot