normal horse serum blocking solution  (Vector Laboratories)

Journal: Poultry Science

Article Title: Double-negative feedback loop between IGF2BP2 and miR-196-5p inhibits the proliferation and differentiation of primary chicken myoblasts

doi: 10.1016/j.psj.2026.106746

Figure Lengend Snippet: The Effect of IGF2BP2 on the Proliferation and Differentiation of Myoblasts. (A) CCK-8 assay assessing cell proliferation following transfection with pcDNA3.1-IGF2BP2 or the negative control pcDNA3.1 at 0, 24, 48, 72, and 96 h (mean ± SEM; within each time point, significance between experimental group and control group was denoted as * P < 0.05; ** P < 0.01; n = 4). (B) EdU assay performed 48 h post-transfection to evaluate cell proliferation (20 ×, scale 50 µm) (mean ± SEM; * P < 0.05; ** P < 0.01; n = 4). (C) The relative mRNA expression levels of IGF2BP2 and proliferation markers genes ( PCNA, CCND1 , and CDK2 ) 48 h after transfection (mean ± SEM; * P < 0.05; ** P < 0.01; n = 4). (D) Relative protein levels of proliferation markers ( PCNA and CDK2 ) 48 h after transfection (mean ± SEM; * P < 0.05; ** P < 0.01; n = 3). (E) Immunofluorescence staining with MyHC (200 × magnification) after transfection. Cells were stained for MyHC in red serving as myogenic marker and DAPI in blue represents the nucleus. The right graph quantifies the relative myotube area (mean ± SEM; * P < 0.05; ** P < 0.01; n = 4). (F) Relative mRNA expression levels of IGF2BP2 and myogenic differentiation markers, including MyHC, MyoD1, MyoG , and Myomaker after 3 days of differentiation induction (mean ± SEM; * P < 0.05; ** P < 0.01; n = 4). (G) Relative protein levels of myogenic differentiation markers (MyHC, MyoD1) post-transfection (mean ± SEM; * P < 0.05; ** P < 0.01; n = 3).

Article Snippet: The cells were then blocked with immunofluorescence blocking solution (Sangon Biotechnology, Shanghai, China) for 1 h at room temperature.

Techniques: CCK-8 Assay, Transfection, Negative Control, Control, EdU Assay, Expressing, Immunofluorescence, Staining, Marker, Cell Characterization

Journal: Poultry Science

Article Title: Double-negative feedback loop between IGF2BP2 and miR-196-5p inhibits the proliferation and differentiation of primary chicken myoblasts

doi: 10.1016/j.psj.2026.106746

Figure Lengend Snippet: IGF2BP2 and miR-196-5p regulate chicken myoblasts differentiation. (A) Immunofluorescence analysis of MyHC cells after co-transfection (200x magnification) (mean ± SEM; n = 4). a, b Among different groups, values with different lowercase superscripts denote significant differences ( P < 0.05). (B) Relative mRNA expression of the myogenic differentiation markers (mean ± SEM; n = 4). a, b Among different groups, values with different lowercase superscripts denote significant differences ( P < 0.05). (C) Protein relative expression levels of myogenic differentiation markers. (mean ± SEM; n = 2). a, b Among different groups, values with different lowercase superscripts denote significant differences ( P < 0.05).

Article Snippet: The cells were then blocked with immunofluorescence blocking solution (Sangon Biotechnology, Shanghai, China) for 1 h at room temperature.

Techniques: Immunofluorescence, Cotransfection, Expressing, Cell Characterization