universal sybr green master mix  (Bio-Rad)

Journal: Journal of Advanced Research

Article Title: JAM-C prevents ocular fibrosis by suppressing the TAZ/KLF6 pathway

doi: 10.1016/j.jare.2025.05.037

Figure Lengend Snippet: TAZ increases KLF6′s expression and pro-EMT function. (A) Volcano plot showing the up- and down-regulated genes in TAZ overexpressing RPE cells. (B) qRT-PCR showing KLF6 level in TAZ overexpressing RPE cells. n = 3. (C-E) Western blot showing the protein levels of TAZ and KLF6 after TAZ knockdown in primary human RPE cells. n = 3. (F) Dual luciferase reporter gene assay showing the luciferase activity of FN1 promoter after the overexpression of TAZ and/or KLF6 in RPE cells. n = 3. (G) ChIP-qPCR assay detected the enrichment of KLF6 at FN1 promoter region in RPE cells. n = 3. (H) qRT-PCR results showing the expression level of FN1 in RPE cells overexpressing KLF6 and/or TAZ. n = 3. (I) qRT-PCR result showing KLF6 mRNA level after knockdown of JAM-C and/or TAZ in RPE cells. n = 4. (J-N) Western blot showing the protein levels of JAM-C, TAZ, KLF6 and fibronectin after the knockdown of JAM-C and/or TAZ in RPE cells. n = 3. Data are presented as mean ± SD. Statistical significance was determined using Student’s t-test in B and one-way ANOVA in D-I, K-N. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: 1 μg RNA was reverse transcribed into cDNA using a cDNA synthesis kit (KR116, TIANGEN, Beijing, China). qRT-PCR was performed using the qPCR SYBR® green master mix (Q711, Vazyme, China) and an ABI QuantStudio 6 Flex device (Life Technologies, Gaithersburg, MD, USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Knockdown, Luciferase, Reporter Gene Assay, Activity Assay, Over Expression, ChIP-qPCR