Journal: Frontiers in Oncology

Article Title: SPAK Deficiency Attenuates Chemotherapy-Induced Intestinal Mucositis

doi: 10.3389/fonc.2021.733555

Figure Lengend Snippet: (A, B) Cell permeability in murine epithelial IEC-6 cells following treatment of 5-FU. (A) After 5-FU treatment, permeability of IEC-6 cells increases in a concentration-dependent manner (5 and 10 μM). (B) siRNA-mediated knockdown of SPAK expression in IEC-6 cells blocks this increase in cell permeability compared to control cells treated with 5-FU (10 μM). (C, D) SPAK-knockdown modulates the effects of 5-FU treatment on the proliferation and survival of IEC-6 cells. Control and SPAK-KD IEC-6 cells were treated with 10 µM 5-FU and dimethyl sulfoxide (DMSO) solvent for 3 h, respectively. Untreated and 5-FU-treated control or SPAK-KD IEC-6 cells were analyzed to determine the mean fluorescence index (MFI) of PCNA expression (C) and the percentage of annexin-V-positive cells (D) , respectively. (C) Representative flow cytometry assays are shown in the left panel, and the MFI of PCNA is shown in the right panel. (D) Representative flow cytometry assays (left panel) and the frequencies (right panel) of annexin-V-positive IEC-6 cells are presented. Data corresponding to MFIs or frequencies are representatives of at least three mice in each group, presented as the mean ± SEM. Two-tailed Student’s unpaired t -tests were used for statistical analysis. *** P < 0.0001; ** P < 0.01; * P < 0.05 n.s., not significant.

Article Snippet: Murine intestinal epithelial IEC-6 cells (Bioresource Collection and Research Center, Hsinchu, Taiwan) were cultured in Dulbecco’s modified Eagle’s medium containing 5% fetal bovine serum, 1.5 g/L sodium bicarbonate, 4.5 g/L glucose, and 10 mM sodium pyruvate supplemented with 0.1 U/mL bovine insulin, as described previously ( ).

Techniques: Permeability, Concentration Assay, Knockdown, Expressing, Control, Solvent, Fluorescence, Flow Cytometry, Two Tailed Test

Journal: Molecular Medicine Reports

Article Title: Atractylenolide III inhibits epithelial-mesenchymal transition in small intestine epithelial cells by activating the AMPK signaling pathway

doi: 10.3892/mmr.2022.12614

Figure Lengend Snippet: Effect of ATL-III on the activity of IEC-6 cells. (A) Chemical structure of ATL-III. (B) Effect of ATL-III on the activity of IEC-6 cells was detected using an MTT assay. ATL-III, atractylenolide III.

Article Snippet: IEC-6 (rat small intestinal epithelial) cells were purchased from Merck-KGaA.

Techniques: Activity Assay, MTT Assay

Journal: Molecular Medicine Reports

Article Title: Atractylenolide III inhibits epithelial-mesenchymal transition in small intestine epithelial cells by activating the AMPK signaling pathway

doi: 10.3892/mmr.2022.12614

Figure Lengend Snippet: ATL-III inhibits the invasion and migration of IEC-6 cells induced by TGF-β1. (A) Cell morphology after TGF-β1 treatment was observed under a microscope. (B) Cell morphology after ATL-III treatment was observed under a microscope. Magnification, ×200. (C) Migration of IEC-6 cells was detected using a wound healing assay. (D) Cell invasion was detected using a Transwell assay. Histograms of cell (E) migration and (F) invasion rates. (G) Expression level of MMP9, as detected via western blotting. Magnification, ×100. ***P<0.001 vs. control group; ## P<0.01, ### P<0.001 vs. TGF-β1 group; n≥3. ATL-III, atractylenolide III.

Article Snippet: IEC-6 (rat small intestinal epithelial) cells were purchased from Merck-KGaA.

Techniques: Migration, Microscopy, Wound Healing Assay, Transwell Assay, Expressing, Western Blot, Control

Journal: Molecular Medicine Reports

Article Title: Atractylenolide III inhibits epithelial-mesenchymal transition in small intestine epithelial cells by activating the AMPK signaling pathway

doi: 10.3892/mmr.2022.12614

Figure Lengend Snippet: ATL-III inhibits the EMT of IEC-6 cells induced by TGF-β1. (A) Expression levels of protein associated with EMT, as detected via western blotting. (B) Expression levels of N-cadherin and (C) E-cadherin were detected using an immunofluorescence assay. Magnification, ×200. ***P<0.001 vs. control group; ## P<0.01, ### P<0.001 vs. TGF-β1 group; n≥3. ATL-III, atractylenolide III; EMT, epithelial-mesenchymal transition; ZO-1, zonula occludens-1.

Article Snippet: IEC-6 (rat small intestinal epithelial) cells were purchased from Merck-KGaA.

Techniques: Expressing, Western Blot, Immunofluorescence, Control

Journal: Molecular Medicine Reports

Article Title: Atractylenolide III inhibits epithelial-mesenchymal transition in small intestine epithelial cells by activating the AMPK signaling pathway

doi: 10.3892/mmr.2022.12614

Figure Lengend Snippet: ATL-III activates the AMPK signaling pathway in IEC-6 cells. (A) Expression levels of p-AMPK and AMPK in the groups treated with different concentrations of ATL-III were measured using western blotting. (B) Expression levels of p-AMPK and AMPK in the groups treated with TGF-β1 and different concentrations of ATL-III were measured using western blotting. (C) Chemical structure of compound C. (D) Expression levels of p-AMPK and AMPK in the groups treated with TGF-β1, ATL-III and compound C were detected using western blotting. *P<0.05, ***P<0.001 vs. control group; ### P<0.001 vs. TGF-β1 group; △△P<0.01 vs. TGF-β1 + ATL-III (20 µmol/l) group; n≥3. ATL-III, atractylenolide III; AMPK, AMP-activated protein kinase; p-, phosphorylated; t-, total.

Article Snippet: IEC-6 (rat small intestinal epithelial) cells were purchased from Merck-KGaA.

Techniques: Expressing, Western Blot, Control

Journal: Molecular Medicine Reports

Article Title: Atractylenolide III inhibits epithelial-mesenchymal transition in small intestine epithelial cells by activating the AMPK signaling pathway

doi: 10.3892/mmr.2022.12614

Figure Lengend Snippet: ATL-III inhibits the invasion, migration and EMT process of IEC-6 cells induced by TGF-β1 by activating the AMPK signaling pathway. (A) Cell migration, as determined using a wound healing assay. (B) Cells invasion was detected using a Transwell assay. Magnification, ×100. Histograms of cell (C) migration and (D) invasion rates. (E) Expression levels of MMP9 and proteins associated with EMT, as detected via western blotting. (F) N-cadherin and (G) E-cadherin expression was detected using an immunofluorescence assay. Magnification, ×200. ***P<0.001 vs. control group; ### P<0.001 vs. TGF-β1 group; △P<0.05, △△P<0.01 vs. TGF-β1 + ATL-III (20 µmol/l) group; n≥3. ATL-III, atractylenolide III; EMT, epithelial-mesenchymal transition; AMPK, AMP-activated protein kinase; ZO-1, zonula occludens-1.

Article Snippet: IEC-6 (rat small intestinal epithelial) cells were purchased from Merck-KGaA.

Techniques: Migration, Wound Healing Assay, Transwell Assay, Expressing, Western Blot, Immunofluorescence, Control