Journal: Molecular Medicine Reports

Article Title: MicroRNA-375 prevents TGF-β-dependent transdifferentiation of lung fibroblasts via the MAP2K6/P38 pathway

doi: 10.3892/mmr.2020.11261

Figure Lengend Snippet: miR-375 suppresses P38 activation by directly inhibiting MAP2K6. (A) Putative binding site of miR-375 in the 3′-UTR of Map2k6 . (B) Statistical analysis of the luciferase reporter assay with WT or MUT 3′-UTR of Map2k6 (n=8). (C and D) Representative western blot and statistical analysis (n=6). (E) Transfection efficiency of si Map2k6 in human lung fibroblasts (n=6). (F) Relative mRNA expression levels of fibrotic markers in human lung fibroblasts transfected with miR-375 inhibitor or si Map2k6 (n=6). Results are expressed as the mean ± SD. *P<0.05. NS no significance; miR , microRNA; MControl, mimic control; IControl, inhibitor control; UTR, untranslated region; WT, wild type; MUT, mutant; si, small interfering RNA; MAP2K6, mitogen-activated protein kinase 6; TGF-β, transforming growth factor-β; Ctgf , connective tissue growth factor; Col1α1 , collagen type I α1; Col3α1 , collagen type III α1; Fn , fibronectin.

Article Snippet: The small interfering RNA (siRNA) against Map2k6 (si Mapk2k6 ; cat. no. sc-35913) was purchased from Santa Cruz Biotechnology, Inc. and scramble control RNA (siRNA, cat. no. sc-37007) was used as the control.

Techniques: Activation Assay, Binding Assay, Luciferase, Reporter Assay, Western Blot, Transfection, Expressing, Control, Mutagenesis, Small Interfering RNA

Journal: Oncogenesis

Article Title: MLK4β functions as a negative regulator of MAPK signaling and cell invasion

doi: 10.1038/oncsis.2012.6

Figure Lengend Snippet: MLK4β negatively regulates p38 and MEK3/MEK6 activation. ( a ) HCT116 cells were transfected with nonspecific or MLK4 siRNA. The cells were left untreated (left panel) or treated (right panel) with 0.5 M sorbitol for 30 min and cell lysates were immunoblotted with MLK4β, p-p38, p38 and β-Actin antibodies. ( b ) HCT116 cells were transiently transfected with pCMV5-FLAG or FLAG-MLK4β and left untreated or treated with 0.5 M sorbitol for 30 min. Cell lysates were immunoblotted with FLAG, p38, p-p38 and actin antibodies. ( c ) HCT116 cells were transfected as described in b and left untreated or treated with 10 μM TNF-α for 20 min. Cell lysates were immunoblotted as described in b . HCT116 ( d ) and HEY1B ( e ) cells were transfected with nonspecific or MLK4 siRNA oligos. Cell lysates were prepared and immunoblotted with MLK4β, p-p38, p38, p-MEK3/MEK6, MEK3/MEK6 and β-Actin antibodies.

Article Snippet: Anti-MEK6 antibody was from Stressgen Bioreagents corporation (Victoria, British Columbia, Canada).

Techniques: Activation Assay, Transfection