Journal: Andrology

Article Title: Identification of MEF2A, MEF2C, and MEF2D interactomes in basal and Fsk-stimulated mouse MA-10 Leydig cells.

doi: 10.1111/andr.70051

Figure Lengend Snippet: FIGURE 1 Characterization of MEF2‒TurboID constructs and biotinylation analysis. (A) Schematic representation of the TurboID constructs, each with a 3xHA tag and a nuclear localization signal (NLS). MEF2‒TurboID fusions have MEF2 positioned at either the N- or C-terminal region of TurboID. TurboID‒GFP and empty TurboID were used as controls. (B) Western blot of 10 µg of nuclear extracts from MA-10 Leydig cells transfected with MEF2A-, MEF2C-, or MEF2D‒TurboID constructs or empty expression vector (Ctr). HA-tag detection confirmed the expression of MEF2A and MEF2D fused to the C-terminal of TurboID (C-MEF2A and C-MEF2D, 100 kDa), and MEF2C to the N-terminal (N-MEF2C, 85 kDa). LaminB was used as a loading control. (C) Reporter assay in MA-10 Leydig cells (left) and CV-1 fibroblast cells (right). Cells were co-transfected with 3xMEF2-luc reporter (pcDNA Ctr), wild-type (WT) MEF2 factors, or MEF2‒TurboID factors (N or C), with or without ERK5 (+ or −). Asterisks (*) indicate a significant difference (p < 0.05) between ERK5 alone (control, white hashed bar) and ERK5 + MEF2 constructs (MEF2‒TurboID, either N- or C-terminal, and WT, colorful hashed bars). Hashtags (#) indicate a significant difference (p < 0.05) between MEF2 alone (colorful solid bar) and ERK5 + MEF2 constructs (colorful hashed bars). Results shown are representative of four independent experiments. (D) Western blot analysis of 30 µg of total protein extracts from MA-10 Leydig cells transfected with C-MEF2A‒TurboID (C-terminal), and either left untreated (−) or treated (+) with 10 µM of forskolin (Fsk) and/or 50 µM of biotin for 4 h. GAPDH was used as a loading control. (E) Western blot of biotinylated proteins in MA-10 Leydig cells transfected with C-MEF2A‒TurboID, TurboID‒MEF2C-N, and C-MEF2D‒TurboID. Blot includes an input 4 h sample (before streptavidin bead enrichment) and biotinylated proteins eluted from streptavidin beads after 1 or 4 h of biotin supplementation, as well as a no-biotin control (−). Streptavidin‒HRP was used to visualize biotinylation.

Article Snippet: Expression vectors for ERK5 (pcDNA3-HA-ERK5) (Addgene plasmid #65244, http://n2t.net/addgene:65244, RRID:Addgene_65244) and the constitutively active MEK5 (pcDNA3-MEK5DD-HA) (Addgene plasmid #65247, http://n2t.net/addgene:65247, RRID:Addgene_65247) were gifts from Dr. Axel Ullrich.22 The 3xHATurboID-NLS_pcDNA3 was a gift from Alice Ting (Addgene plasmid #107171, http://n2t.net/addgene:107171, RRID:Addgene_107171).

Techniques: Construct, Western Blot, Transfection, Expressing, Plasmid Preparation, Control, Reporter Assay

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Effects of the Wenyang Zhenshuai Granule on the Expression of LncRNA-MiR143HG/miR-143 Regulating ERK5 in H9C2 Cardiomyocytes

doi: 10.1155/2021/6431007

Figure Lengend Snippet: qRT-PCR primers.

Article Snippet: Adriamycin hydrochloride was provided by Shenzhen Wanle Pharmaceutical Co., Ltd. (H44024359), SYBR Green PCR kit was purchased from Shanghai Sixin Biotechnology Co., Ltd. (BL705A), and Wenyang Zhenshuai granules were purchased from the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine Provided (201902), and ERK5 and p-ERK5 antibodies were purchased from Wuhan Boster Bioengineering Co., Ltd. (ab40908, ab5686).

Techniques: Sequencing

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Effects of the Wenyang Zhenshuai Granule on the Expression of LncRNA-MiR143HG/miR-143 Regulating ERK5 in H9C2 Cardiomyocytes

doi: 10.1155/2021/6431007

Figure Lengend Snippet: Comparison of ERK5 expression in cardiomyocytes of various groups. Note: compared with the normal control group, ∗ P < 0.05; compared with the ADR group, # P < 0.05; compared with the ADR + LncRNA-MiR143HG silence + medicated serogroup, + P < 0.05.

Article Snippet: Adriamycin hydrochloride was provided by Shenzhen Wanle Pharmaceutical Co., Ltd. (H44024359), SYBR Green PCR kit was purchased from Shanghai Sixin Biotechnology Co., Ltd. (BL705A), and Wenyang Zhenshuai granules were purchased from the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine Provided (201902), and ERK5 and p-ERK5 antibodies were purchased from Wuhan Boster Bioengineering Co., Ltd. (ab40908, ab5686).

Techniques: Comparison, Expressing, Control

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Effects of the Wenyang Zhenshuai Granule on the Expression of LncRNA-MiR143HG/miR-143 Regulating ERK5 in H9C2 Cardiomyocytes

doi: 10.1155/2021/6431007

Figure Lengend Snippet: Comparison of the expression of p-ERK5 and ERK5 proteins in cardiomyocytes of each group. (a) Normal control group, (b) LncRNA-MiR143HG overexpression group, (c) LncRNA-MiR143HG silence group, (d) ADR group, (e) ADR + medicated serum group, (f) ADR + LncRNA-MiR143HG overexpression + medicated serogroup, and (g) ADR + LncRNA-MiR143HG silence + medicated serogroup. Compared with the normal control group, ∗ P < 0.05; compared with the ADR group, # P < 0.05; compared with the ADR + LncRNA-MiR143HG silence + medicated serogroup, + P < 0.05.

Article Snippet: Adriamycin hydrochloride was provided by Shenzhen Wanle Pharmaceutical Co., Ltd. (H44024359), SYBR Green PCR kit was purchased from Shanghai Sixin Biotechnology Co., Ltd. (BL705A), and Wenyang Zhenshuai granules were purchased from the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine Provided (201902), and ERK5 and p-ERK5 antibodies were purchased from Wuhan Boster Bioengineering Co., Ltd. (ab40908, ab5686).

Techniques: Comparison, Expressing, Control, Over Expression