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Sangon Biotech mettl3 coding sequence
( A) Details of seven potential methylation modification sites on circSIK2. (B ) The relative expression of circSIK2, <t>METTL3,</t> and SIK2 mRNA in myoblast cells after transfected with pCD25-circSIK2 + pCD3.1, pCD25-circSIK2 + pCD3.1-METTL3, and pCD25 + pCD3.1. ( C) Upper: the expression of SIK2-176aa-flag after transfected with pCD3.1-SIK2-176aa-flag and pCD3.1-SIK2-176aa-flag + pCD3.1-METTL3. Lower: the expression of SIK2-176aa after transfected with pCD3.1-SIK2-176aa-flag, pCD25-circSIK2, and pCD25-circSIK2 + pCD3.1-METTL3. (D ) Dual-luciferase reporter assay using the different regions of the circSIK2 after co-transfected with METTL3 in DF-1 cells. ( E) Upper: Dual-luciferase reporter assay using the 151–200 fragment after co-transfected with METTL3 in DF-1 cells. Under: Dual-luciferase reporter assay after 131A-WT and 131A-MT fragment was co-transfected with METTL14 in DF-1 cells. (F ) Amplification curve and qPCR CT value in circSIK2 131 m 6 A site and 125 A site after METTL3 overexpression in myoblast cells. ( G, H) circSIK2 pulldown image ( G ) (OV = pCD25-circSIK2, NC = pCD25) and binding proteins’ string network ( H ) screened by mass spectrometry in myoblast cells. ( I ) circSIK2 pulldown results confirmed by western blot with HNRNPA2B1 antibody in myoblast cells. (J) The expression pattern of METTL3, METTL14, EIF3A, and EIF3F in different stages of myoblast differentiation. GM (growth media; n = 3) stands for myoblasts in the proliferative phase. DM (differentiation media; n = 3), DM1-DM6 means differentiation from day 1 to day 6.
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1) Product Images from "METTL3 facilitates the translation of CircSIK2 during chicken myogenesis in an m 6 A dependent manner"

Article Title: METTL3 facilitates the translation of CircSIK2 during chicken myogenesis in an m 6 A dependent manner

Journal: PLOS Genetics

doi: 10.1371/journal.pgen.1011934

( A) Details of seven potential methylation modification sites on circSIK2. (B ) The relative expression of circSIK2, METTL3, and SIK2 mRNA in myoblast cells after transfected with pCD25-circSIK2 + pCD3.1, pCD25-circSIK2 + pCD3.1-METTL3, and pCD25 + pCD3.1. ( C) Upper: the expression of SIK2-176aa-flag after transfected with pCD3.1-SIK2-176aa-flag and pCD3.1-SIK2-176aa-flag + pCD3.1-METTL3. Lower: the expression of SIK2-176aa after transfected with pCD3.1-SIK2-176aa-flag, pCD25-circSIK2, and pCD25-circSIK2 + pCD3.1-METTL3. (D ) Dual-luciferase reporter assay using the different regions of the circSIK2 after co-transfected with METTL3 in DF-1 cells. ( E) Upper: Dual-luciferase reporter assay using the 151–200 fragment after co-transfected with METTL3 in DF-1 cells. Under: Dual-luciferase reporter assay after 131A-WT and 131A-MT fragment was co-transfected with METTL14 in DF-1 cells. (F ) Amplification curve and qPCR CT value in circSIK2 131 m 6 A site and 125 A site after METTL3 overexpression in myoblast cells. ( G, H) circSIK2 pulldown image ( G ) (OV = pCD25-circSIK2, NC = pCD25) and binding proteins’ string network ( H ) screened by mass spectrometry in myoblast cells. ( I ) circSIK2 pulldown results confirmed by western blot with HNRNPA2B1 antibody in myoblast cells. (J) The expression pattern of METTL3, METTL14, EIF3A, and EIF3F in different stages of myoblast differentiation. GM (growth media; n = 3) stands for myoblasts in the proliferative phase. DM (differentiation media; n = 3), DM1-DM6 means differentiation from day 1 to day 6.
Figure Legend Snippet: ( A) Details of seven potential methylation modification sites on circSIK2. (B ) The relative expression of circSIK2, METTL3, and SIK2 mRNA in myoblast cells after transfected with pCD25-circSIK2 + pCD3.1, pCD25-circSIK2 + pCD3.1-METTL3, and pCD25 + pCD3.1. ( C) Upper: the expression of SIK2-176aa-flag after transfected with pCD3.1-SIK2-176aa-flag and pCD3.1-SIK2-176aa-flag + pCD3.1-METTL3. Lower: the expression of SIK2-176aa after transfected with pCD3.1-SIK2-176aa-flag, pCD25-circSIK2, and pCD25-circSIK2 + pCD3.1-METTL3. (D ) Dual-luciferase reporter assay using the different regions of the circSIK2 after co-transfected with METTL3 in DF-1 cells. ( E) Upper: Dual-luciferase reporter assay using the 151–200 fragment after co-transfected with METTL3 in DF-1 cells. Under: Dual-luciferase reporter assay after 131A-WT and 131A-MT fragment was co-transfected with METTL14 in DF-1 cells. (F ) Amplification curve and qPCR CT value in circSIK2 131 m 6 A site and 125 A site after METTL3 overexpression in myoblast cells. ( G, H) circSIK2 pulldown image ( G ) (OV = pCD25-circSIK2, NC = pCD25) and binding proteins’ string network ( H ) screened by mass spectrometry in myoblast cells. ( I ) circSIK2 pulldown results confirmed by western blot with HNRNPA2B1 antibody in myoblast cells. (J) The expression pattern of METTL3, METTL14, EIF3A, and EIF3F in different stages of myoblast differentiation. GM (growth media; n = 3) stands for myoblasts in the proliferative phase. DM (differentiation media; n = 3), DM1-DM6 means differentiation from day 1 to day 6.

Techniques Used: Methylation, Modification, Expressing, Transfection, Luciferase, Reporter Assay, Amplification, Over Expression, Binding Assay, Mass Spectrometry, Western Blot



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