Journal: JID Innovations
Article Title: Targeting TGF-β1/miR-21 Pathway in Keratinocytes Reveals Protective Effects of Silymarin on Imiquimod-Induced Psoriasis Mouse Model
doi: 10.1016/j.xjidi.2022.100175
Figure Lengend Snippet: SM positively regulates miR-21-5p through a SMAD-independent signaling pathway in keratinocytes. ( a ) Quantitative bioluminescence values were detected in indicated HaCaT cell groups and expressed as luciferase fold induction relative to control cells transfected with the control pRILES. ( b ) Western blot analysis of SMAD2, SMAD3, and P-SMAD2 protein levels in total cell lysates from HaCaT cells treated with 10 ng/ml TGF-β1 in the presence or absence of indicated concentrations of SM for 1 h. β-Actin was used as a loading control. Band intensities were quantitatively analyzed using the ImageJ software. Bar graph data represent the ratio of protein of interest/β-actin. ( c ) RT-qPCR quantification of the relative expression of FN, MMP3, and CDH1 to GAPDH levels in HaCaT cells treated with 10 ng/ml TGF-β1 in the presence or absence of indicated concentrations of SM for 24 h. Untreated cells were used as the negative control. ( d ) In vitro scratch assay assessing the migration rate of HaCaT cells treated with 10 ng/ml TGF-β1 in the presence or absence of SM (31 μg/ml) for 48 h. Untreated cells were used as the negative control. The wound closure area was calculated using Zen 2.3 SP1 (Carl Zeiss, White Plains, NY). Data are presented as mean ± SEM and are representative of three independent experiments performed in triplicate. ANOVA test was applied to test for significance between the different groups in a , b, and c. Repeated measures ANOVA tests were used for d . ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001. FN, fibronectin; h, hour; MMP3, metalloproteinase-3; ns, not significant; P-SMAD2, phosphorylated SMAD2; SM, silymarin.
Article Snippet: Wound areas at individual time points were analyzed using the ZEN LE Digital Imaging Software (Carl Zeiss).
Techniques: Luciferase, Control, Transfection, Western Blot, Software, Quantitative RT-PCR, Expressing, Negative Control, In Vitro, Wound Healing Assay, Migration
Carl Zeiss is a verified supplier 
Carl Zeiss manufactures this product 
