Journal: Hypertension

Article Title: Transcribed Ultraconserved Regions, Uc.323, Ameliorates Cardiac Hypertrophy by Regulating the Transcription of CPT1b (Carnitine Palmitoyl transferase 1b)

doi: 10.1161/hypertensionaha.119.13173

Figure Lengend Snippet: Figure 4. Effects of uc.323 on the ERK1/2, mTORC1, and phosphorylated ribosomal protein S6 kinase signaling pathways. A, Western blots and corresponding quantitative analysis showing the effects of uc.323 knockdown on the ERK1/2, mTORC1, and p70S6K signaling pathways; n=5–6. B, Western blots and corresponding quantitative analysis showing the effects of uc.323 overexpression on the ERK1/2, mTORC1, and p70S6K signaling pathways; n=4–5. C, quantitative real-time polymerase chain reaction analysis showed that rapamycin could block downregulation of uc.323 by phenylephrine (PE); n=4. D, Effects of uc.323 knockdown and rapamycin treatment on the mRNA expression of ANP (atrial natriuretic peptide) and BNP (B-type natriuretic peptide); GAPDH was used as an internal control; n=5. E, Effects of uc.323 knockdown and rapamycin treatment on cell cross-sectional area; n=3. Cardiomyocytes were stained with troponin I, and the nuclei were stained with DAPI. NS denotes no significant difference vs the corresponding control group. LV indicates left ventricular; p-ERK1/2, phosphorylated extracellular regulated protein kinases1/2; p-mTORC1, phosphorylated mammalian target of rapamycin 1; si-scramble, scrambled siRNA; and si-uc.323, siRNA targeting uc.323. *P<0.05 vs the corresponding control group. **P<0.01 vs the corresponding control group.

Article Snippet: Monoclonal antibodies against extracellular regulated protein kinases 1/2 (No. 8544S), phospho-ERK1/2 (No. 4695S), c-Jun N-terminal kinase 1/2 (No. 9252), phospho-JNK1/2 (No. 4668), p38 (No. 8690), phospho-p38 (No. 4511), mTORC1 (mammalian target of rapamycin 1) (No. 2587), phospho-mTORC1 (No. 5536), p70S6K (ribosomal protein S6 kinase; No. 2708), phospho-p70S6K (No. 9204), EZH2 (enhancer of zeste homolog 2; No. 5246), H3K27me3 (trimethylation of lysine 27 of histone H3; No. 9733), and normal rabbit IgG (No. 2729) were purchased from Cell Signaling Technology (MA).

Techniques: Protein-Protein interactions, Western Blot, Knockdown, Over Expression, Real-time Polymerase Chain Reaction, Blocking Assay, Expressing, Control, Staining

Journal: Oncotarget

Article Title: Lactobacillus reuteri ZJ617 maintains intestinal integrity via regulating tight junction, autophagy and apoptosis in mice challenged with lipopolysaccharide

doi: 10.18632/oncotarget.20536

Figure Lengend Snippet: Mice were orally inoculated with lactobacilli for 7 days before the challenge with an intraperitoneal injection of 10 mg/kg LPS. Ileal tissues from six mice in each group were obtained 24 h after injection. Immunoblotting of and quantitative analysis of ileal mammalian target of rapamycin (mTOR). (A) Immunoblotting of and quantitative analysis of ileal mTOR. The protein bands were quantified by densitometry analysis, normalized to GAPDH. The values are expressed as means ± SD (n = 6). (B) Immunohistochemistry of and quantitative analysis of ileal mTOR. Brown staining indicates mTOR positive cells. The ratio of positive-stained cells/all cells is expressed as the mean ± SD (n= 6). Different alphabet indicates significance ( P < 0.05).

Article Snippet: The blotted membrane was blocked for 2 h at room temperature in 1 × TBST [0.05% Tween 20, 100 mM Tris–HCl and 150 mM NaCl (pH 7.5)] containing 5% fat-free dry milk, and then incubated under gentle agitation all the night at room temperature in the presence of the primary antibodies: occludin, 1:5000 dilution of purified rabbit polyclonal anti-occludin antibody (Abcam, AB31721, Cambridge, UK); claudin-3, 1: 5000 dilution of purified rabbit polyclonal anti- claudin-3 antibody (Abcam, AB15102, Cambridge, UK); caspase-3, 1:5000 dilution of purified rabbit polyclonal anti- caspase-3 antibody (Abcam, AB13847, Cambridge, UK); glyceraldehyde-3-phophate dehydrogenase (GAPDH), 1:1000 dilution of purified rabbit monoclonal anti-GAPDH antibody (CST, D16H11, Danvers, USA); LC3, 1:1000 dilution of purified rabbit monoclonal anti-LC3 protein antibody (CST, D3U4C, Danvers, USA); mTOR, 1:1000 dilution of purified rabbit monoclonal anti-mTOR protein antibody (CST, 7C10, Danvers, USA); p-mTOR, 1:1000 dilution of purified rabbit monoclonal anti- p-mTOR protein antibody (CST, D3F9, Danvers, USA), which were able to bind to their specific protein.

Techniques: Injection, Western Blot, Immunohistochemistry, Staining