Journal: BioMed Research International

Article Title: Molecular Hydrogen Inhibits Colorectal Cancer Growth via the AKT/SCD1 Signaling Pathway

doi: 10.1155/2022/8024452

Figure Lengend Snippet: Hydrogen inhibits CRC cell proliferation by decreasing pAKT/SCD1. (a) IHC assay in tumors (RKO cells) after H 2 treatment and control (100x). (b) Western blot analysis showing the expression levels of p-PI3K, PI3K, AKT, pAKT, and SCD1 in RKO, SW480, and HCT116 cells before and after H 2 treatment. Western blotting assays showed that the expression of SCD1 was significantly upregulated and presented a dose-dependent effect with SC79 treatment. (c) The inhibition of cell proliferation induced by H 2 was reversed when cells were treated with SC79, as determined by the CCK-8 assay.

Article Snippet: Total proteins from cells (20 μ g) and tissues (20 μ g) were separated by SDS PAGE through a 10% gel and transferred to a polyvinylidene fluoride (PVDF) membrane (Millipore, IPVH00010, MA, USA) at 100 V for 60 min. After blocked with Tris-buffered saline containing Tween-20 (TBST, 1000 : 1) and 5% fat-free milk for 2 h, the membranes were incubated at 4°C overnight with primary antibodies against SCD1 (1 : 1000; Bioss, Cat# bs-3787R, Beijing, China), p-PI3K (1 : 1000; Cat#: 17366; Cell Signaling Technology, Danvers, MA, USA), PI3K (1 : 1000; clone: 1F6A7; Proteintech, Wuhan, China), phospho-AKT (Ser473; 1 : 1000; Cat# 4069; Cell Signaling Technology, Danvers, MA, USA), AKT (1 : 1000; Cat# 4691; Cell Signaling Technology), and actin (rabbit polyclonal; 1 : 2000; Cat# E-AB-20058; Elabscience, Wuhan, China).

Techniques: Western Blot, Expressing, Inhibition, CCK-8 Assay

Journal: BioMed Research International

Article Title: Molecular Hydrogen Inhibits Colorectal Cancer Growth via the AKT/SCD1 Signaling Pathway

doi: 10.1155/2022/8024452

Figure Lengend Snippet: The expression of SCD1 in paraffin-embedded CRC and normal epithelium tissues. SCD1 expression levels in malignant cells were significantly higher than those in normal epithelial tissues (100x).

Article Snippet: Total proteins from cells (20 μ g) and tissues (20 μ g) were separated by SDS PAGE through a 10% gel and transferred to a polyvinylidene fluoride (PVDF) membrane (Millipore, IPVH00010, MA, USA) at 100 V for 60 min. After blocked with Tris-buffered saline containing Tween-20 (TBST, 1000 : 1) and 5% fat-free milk for 2 h, the membranes were incubated at 4°C overnight with primary antibodies against SCD1 (1 : 1000; Bioss, Cat# bs-3787R, Beijing, China), p-PI3K (1 : 1000; Cat#: 17366; Cell Signaling Technology, Danvers, MA, USA), PI3K (1 : 1000; clone: 1F6A7; Proteintech, Wuhan, China), phospho-AKT (Ser473; 1 : 1000; Cat# 4069; Cell Signaling Technology, Danvers, MA, USA), AKT (1 : 1000; Cat# 4691; Cell Signaling Technology), and actin (rabbit polyclonal; 1 : 2000; Cat# E-AB-20058; Elabscience, Wuhan, China).

Techniques: Expressing

Journal: BioMed Research International

Article Title: Molecular Hydrogen Inhibits Colorectal Cancer Growth via the AKT/SCD1 Signaling Pathway

doi: 10.1155/2022/8024452

Figure Lengend Snippet: SCD1 status and its association with clinicopathological characteristics in CRC patients.

Article Snippet: Total proteins from cells (20 μ g) and tissues (20 μ g) were separated by SDS PAGE through a 10% gel and transferred to a polyvinylidene fluoride (PVDF) membrane (Millipore, IPVH00010, MA, USA) at 100 V for 60 min. After blocked with Tris-buffered saline containing Tween-20 (TBST, 1000 : 1) and 5% fat-free milk for 2 h, the membranes were incubated at 4°C overnight with primary antibodies against SCD1 (1 : 1000; Bioss, Cat# bs-3787R, Beijing, China), p-PI3K (1 : 1000; Cat#: 17366; Cell Signaling Technology, Danvers, MA, USA), PI3K (1 : 1000; clone: 1F6A7; Proteintech, Wuhan, China), phospho-AKT (Ser473; 1 : 1000; Cat# 4069; Cell Signaling Technology, Danvers, MA, USA), AKT (1 : 1000; Cat# 4691; Cell Signaling Technology), and actin (rabbit polyclonal; 1 : 2000; Cat# E-AB-20058; Elabscience, Wuhan, China).

Techniques: Expressing, Mutagenesis

Journal: BioMed Research International

Article Title: Molecular Hydrogen Inhibits Colorectal Cancer Growth via the AKT/SCD1 Signaling Pathway

doi: 10.1155/2022/8024452

Figure Lengend Snippet: The prognostic value of SCD1 expression in primary CRC. Survival curves for disease free survival (DFS) and overall survival (OS) in stages I–III colorectal cancer according to SCD1 status: (a) DFS according to SCD1; (b) OS according to SCD1.

Article Snippet: Total proteins from cells (20 μ g) and tissues (20 μ g) were separated by SDS PAGE through a 10% gel and transferred to a polyvinylidene fluoride (PVDF) membrane (Millipore, IPVH00010, MA, USA) at 100 V for 60 min. After blocked with Tris-buffered saline containing Tween-20 (TBST, 1000 : 1) and 5% fat-free milk for 2 h, the membranes were incubated at 4°C overnight with primary antibodies against SCD1 (1 : 1000; Bioss, Cat# bs-3787R, Beijing, China), p-PI3K (1 : 1000; Cat#: 17366; Cell Signaling Technology, Danvers, MA, USA), PI3K (1 : 1000; clone: 1F6A7; Proteintech, Wuhan, China), phospho-AKT (Ser473; 1 : 1000; Cat# 4069; Cell Signaling Technology, Danvers, MA, USA), AKT (1 : 1000; Cat# 4691; Cell Signaling Technology), and actin (rabbit polyclonal; 1 : 2000; Cat# E-AB-20058; Elabscience, Wuhan, China).

Techniques: Expressing

Journal: Life Metabolism

Article Title: Regulation of hepatic lipid metabolism by intestine epithelium-derived exosomes

doi: 10.1093/lifemeta/load044

Figure Lengend Snippet: miR-21a-5p and miR-145a-5p regulate liver lipid metabolism through direct suppression of Ccl1 and Btg1 , respectively. (a–e) CCL1-dependent mechanism by which miR-21a-5p regulates liver lipid metabolism. (a) Dual-luciferase assay was performed to validate the direct interaction between miR-21a-5p and 3ʹ-UTR of Ccl1 gene. (b) Raw264.7 cells were transfected with miR-21a-5p mimics or scramble sequences, and the mRNA levels of inflammation marker genes were analyzed by RT-qPCR. (c and d) Representative F4/80 staining of liver section and numbers of F4/80 + cells in HFD-induced obese mice injected with AAV9-miR-21a-5p or AAV-9 scramble. (e) Cellular TG and cholesterol in AML12 cells co-cultured with miR-21a-5p-treated Raw264.7 cells. Results were expressed as mean ± SEM and analyzed by one-way or two-way ANOVA, or Student’s t -test, n = 5 − 7, * denotes P < 0.05. (f–k) MiR-145a-5p regulates liver lipid metabolism through direct suppression of Btg1 . (f) Dual-luciferase assay was performed to validate the direct interaction between miR-145a-5p and 3ʹ-UTR of Btg1 gene. (g) TG concentration of AML12 cells transfected with miR-145a-5p mimics and a plasmid expressing Btg1 -encoding sequences without 3ʹ-UTR. Protein expression of SCD1 in livers of mice treated with AAV9-miR-145a-5p-sponge (h and i) or intExo/miR-145a-5p (j and k). Results were expressed as mean ± SEM and analyzed by one-way ANOVA or Student’s t -test, n = 4 − 8, * denotes P < 0.05.

Article Snippet: SCD1 Rabbit pAb , ABclonal , A16429.

Techniques: Luciferase, Transfection, Marker, Quantitative RT-PCR, Staining, Injection, Cell Culture, Concentration Assay, Plasmid Preparation, Expressing

Journal: Life Metabolism

Article Title: Regulation of hepatic lipid metabolism by intestine epithelium-derived exosomes

doi: 10.1093/lifemeta/load044

Figure Lengend Snippet: qPCR primers.

Article Snippet: SCD1 Rabbit pAb , ABclonal , A16429.

Techniques:

Journal: Life Metabolism

Article Title: Regulation of hepatic lipid metabolism by intestine epithelium-derived exosomes

doi: 10.1093/lifemeta/load044

Figure Lengend Snippet:

Article Snippet: SCD1 Rabbit pAb , ABclonal , A16429.

Techniques: