mouse anti wtap mab  (Proteintech)

Journal: Frontiers in Molecular Biosciences

Article Title: Alteration of N6-Methyladenosine RNA Profiles in Cisplatin-Induced Acute Kidney Injury in Mice

doi: 10.3389/fmolb.2021.654465

Figure Lengend Snippet: Sequences of primers used for qPCR and MeRIP-qPCR analysis.

Article Snippet: The membranes were subsequently immersed in 5% bovine serum albumin (BSA) in the presence of the polyclonal rabbit anti-cleaved caspase-3 antibody (1:1000, Cell Signaling Technology, MA, United States), the monoclonal rabbit anti-Mettl14 antibody (1:1000, Cell Signaling Technology, MA, United States), the polyclonal rabbit anti-Mettl3 antibody (1:1000, Proteintech Group, Inc, IL, United States), the monoclonal mouse anti-Wtap antibody (1:1000, Proteintech Group, Inc, IL, United States) or the monoclonal mouse anti-Fto antibody (1:1000, Abcam, United Kingdom), the monoclonal rabbit anti-Alkbh5 antibody (1:1000, Abcam, United Kingdom), and the monoclonal mouse anti-GAPDH antibody (1:10000, CoWin Biosciences, China) overnight at 4°C and hybridized with a horseradish peroxidase (HRP)–labeled goat anti-rabbit/-mouse antibody (1:8000; TransGen Biotech, China) at room temperature for 1 h. The blots were washed with TBST and then visualized by enhanced chemiluminescence (ECL; Millipore, MA, United States) through a luminescent imaging workstation (Tanon, China).

Techniques: Sequencing

Journal: Frontiers in Molecular Biosciences

Article Title: Alteration of N6-Methyladenosine RNA Profiles in Cisplatin-Induced Acute Kidney Injury in Mice

doi: 10.3389/fmolb.2021.654465

Figure Lengend Snippet: Cisplatin increased the total m 6 A level and modulated the m 6 A-related genes in the mouse kidneys. (A) Quantification of m 6 A abundance in total RNA isolated from the kidneys of male mice treated with vehicle or cisplatin. The histogram showing the levels of Mettl3 (B) , Mettl14 (C) , Wtap (D) , Fto (E) , and Alkbh5. (F) mRNA expression was quantified by qPCR analysis. GAPDH was used as a housekeeping gene for normalizing changes in specific gene expressions. * p < 0.05 vs. control group, n = 4 per group. Representative immunostaining images and densitometric analysis were performed to assess the protein expression of m 6 A-related enzymes: Mettl3 (G and H) , Mettl14 (I and J) , Wtap (K and L) , Fto (M and N) , and Alkbh5 (O and P) in kidneys of Cis-AKI groups compared with control groups. GAPDH was used as an internal control. Data were expressed as mean ± SEM and analyzed using Student’s t -test to determine the differences between the two groups. * p < 0.05 vs. control group, n = 4 per group.

Article Snippet: The membranes were subsequently immersed in 5% bovine serum albumin (BSA) in the presence of the polyclonal rabbit anti-cleaved caspase-3 antibody (1:1000, Cell Signaling Technology, MA, United States), the monoclonal rabbit anti-Mettl14 antibody (1:1000, Cell Signaling Technology, MA, United States), the polyclonal rabbit anti-Mettl3 antibody (1:1000, Proteintech Group, Inc, IL, United States), the monoclonal mouse anti-Wtap antibody (1:1000, Proteintech Group, Inc, IL, United States) or the monoclonal mouse anti-Fto antibody (1:1000, Abcam, United Kingdom), the monoclonal rabbit anti-Alkbh5 antibody (1:1000, Abcam, United Kingdom), and the monoclonal mouse anti-GAPDH antibody (1:10000, CoWin Biosciences, China) overnight at 4°C and hybridized with a horseradish peroxidase (HRP)–labeled goat anti-rabbit/-mouse antibody (1:8000; TransGen Biotech, China) at room temperature for 1 h. The blots were washed with TBST and then visualized by enhanced chemiluminescence (ECL; Millipore, MA, United States) through a luminescent imaging workstation (Tanon, China).

Techniques: Isolation, Expressing, Control, Immunostaining