purple loading dye  (New England Biolabs)

Journal: Biochemistry and Biophysics Reports

Article Title: Oroxylin A ameliorates hepatic steatosis and non-alcoholic fatty liver disease through the regulation of CPT1A m 6 A methylation in hepatocytes

doi: 10.1016/j.bbrep.2026.102598

Figure Lengend Snippet: Oroxylin A suppressed de novo lipogenesis (DNL) in PA-induced hepatocytes. (A–D) The mRNA levels of CPT-1A, PPARα, SREBP1-C and FASN in AML-12 cells were measured by quantitative PCR (n = 3). (E) Representative bands of CPT-1A, PPARα, SREBP1-C and FASN in AML-12 cells were examined by Western blotting (n = 3). (F–I) Quantification of protein expression in AML-12 cells (n = 3). PA (0.25 mM) was used in A-I. Data are presented as the mean ± SD. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

Article Snippet: RNA was extracted from liver or AML-12 cell lysates and reverse transcribed into cDNA using HiScript IV All-in-One Ultra RT SuperMix. cDNA was amplified using SupRealQ Purple Universal SYBR qPCR Master Mix (Vazyme).

Techniques: Real-time Polymerase Chain Reaction, Western Blot, Expressing

Journal: Biochemistry and Biophysics Reports

Article Title: Oroxylin A ameliorates hepatic steatosis and non-alcoholic fatty liver disease through the regulation of CPT1A m 6 A methylation in hepatocytes

doi: 10.1016/j.bbrep.2026.102598

Figure Lengend Snippet: Oroxylin A regulated the m 6 A demethylation of CPT1A mRNA in PA-stimulated hepatocytes. (A–F) The mRNA levels of m 6 A demethylation Writers in AML-12 cells were measured by quantitative PCR (n = 3). (G-J) The mRNA levels of m 6 A demethylating erasers in AML-12 cells were measured by quantitative PCR (n = 3). (K, L) The mRNA levels of m 6 A demethylation Readers in AML-12 cells were measured by quantitative PCR (n = 3). (M) m 6 A modification levels in AML-12 cells were measured by m 6 A dot blot hybridization. (N) The possible m 6 A modification sites of CPT1A mRNA were analyzed using an online website. PA (0.25 mM) was used in A-M. Data are presented as the mean ± SD. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

Article Snippet: RNA was extracted from liver or AML-12 cell lysates and reverse transcribed into cDNA using HiScript IV All-in-One Ultra RT SuperMix. cDNA was amplified using SupRealQ Purple Universal SYBR qPCR Master Mix (Vazyme).

Techniques: Real-time Polymerase Chain Reaction, Modification, Dot Blot, Hybridization

Journal: Biochemistry and Biophysics Reports

Article Title: Oroxylin A ameliorates hepatic steatosis and non-alcoholic fatty liver disease through the regulation of CPT1A m 6 A methylation in hepatocytes

doi: 10.1016/j.bbrep.2026.102598

Figure Lengend Snippet: Oroxylin A-regulated YTHDC1-mediated m6A modification of CPT1A contributes to a reduction in lipid metabolism in hepatocytes. YTHDC1 siRNA was used for YTHDC1 silencing. (A) The mRNA levels of YTHDC1 in AML-12 cells were measured by quantitative PCR (n = 3). (B–E) The mRNA levels of CPT-1A, PPARα, SREBP1-C and FASN in AML-12 cells were measured by quantitative PCR (n = 3). (F, G) TC and TG levels in AML-12 cells. (H) Lipid droplet accumulation in AML-12 cells was examined by Oil Red O staining (n = 3). PA (0.25 mM) was used in A-M. Data are presented as the mean ± SD. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

Article Snippet: RNA was extracted from liver or AML-12 cell lysates and reverse transcribed into cDNA using HiScript IV All-in-One Ultra RT SuperMix. cDNA was amplified using SupRealQ Purple Universal SYBR qPCR Master Mix (Vazyme).

Techniques: Modification, Real-time Polymerase Chain Reaction, Staining