Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: CircFBXW4 Suppresses Colorectal Cancer Progression by Regulating the MiR-338-5p/SLC5A7 Axis.

doi: 10.1002/advs.202300129

Figure Lengend Snippet: Figure 7. CircFBXW4 regulates SLC5A7 expression by acting as a sponge for miR-338-5p. A) Relative expression of candidate mRNAs in SW480 cells transfected with the miR-338-5p mimic. B) Relative expression of candidate mRNAs in SW620 cells transfected with the miR-338-5p inhibitor. C) Relative protein levels of SLC5A7 in CRC cells transfected with the miR-338-5p mimic or inhibitor. D) The relative expression of SLC5A7 in CRC tissues and matched adjacent normal tissues was determined by qRT‒PCR (n = 40). E) Pearson correlation analysis between the expression levels of miR-338-5p and SLC5A7 in our own patient cohort (n = 40). F) Schematic illustration of the SLC5A7-WT and SLC5A7-MUT luciferase reporter vectors. G) Relative luciferase activity was measured in 293T cells after cotransfection with SLC5A7-WT or SLC5A7-MUT and the miR-338-5p mimic or NC. H) The relative mRNA and protein expression levels of SLC5A7 were measured by qRT‒PCR and western blotting in SW480 cells transfected with si-NC or si-circFBXW4 with or without the miR-338-5p inhibitor. I) The relative mRNA and protein levels of SLC5A7 were measured by qRT‒PCR and western blotting in SW620 cells transfected with vector or the circFBXW4-OE plasmid with or without the miR-338-5p mimic. The data are shown as the means ± SDs; *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The primary antibodies used to detect Ki-67, PCNA, and Caspase3 were purchased from ABclonal (China), and the primary antibody used to detect SLC5A7 was purchased from ProteinTech (China).

Techniques: Expressing, Transfection, Luciferase, Activity Assay, Cotransfection, Western Blot, Plasmid Preparation

Journal: Brain : a journal of neurology

Article Title: Choline transporter mutations in severe congenital myasthenic syndrome disrupt transporter localization.

doi: 10.1093/brain/awx249

Figure Lengend Snippet: Figure 1 Family pedigrees, and mutations detected in SLC5A7. (A–C) Family pedigrees and whole-exome sequencing results. Whole- exome sequencing was done for affected members of each family (Family 1 Patient IV:1, Family 2 Patient V:3 and Family 3 Patient III:5) revealed a single novel deleterious homozygous variant in each subject in the same gene SLC5A7. For Family 3, two siblings (Patients III:3 and III:4) affected with a separate genetic disorder (Fowler syndrome) are marked out in grey. (D) Sequence alignment displays both affected residues (p.Ser94Arg and p.Val112Glu) high evolutionarily sequence conservation while p.Pro210Leu less conserved. (E) CHT topology. The 580 amino acid CHT protein has 13 transmembrane spanning domains and the N-terminus is extracellular while C-terminus is intracellular. The three amino acid substitutions from the missense mutations are indicated in brackets. Two sets of constructs are used in this study, the tags of each set are marked out at the termini.

Article Snippet: The SLC5A7 cDNA (NM_020815) was purchased from Origene in pCMV6-Entry vector with a myc/DDK tag in the C-terminus.

Techniques: Sequencing, Variant Assay, Construct

Journal: PLoS ONE

Article Title: A Non-Neuronal Cardiac Cholinergic System Plays a Protective Role in Myocardium Salvage during Ischemic Insults

doi: 10.1371/journal.pone.0050761

Figure Lengend Snippet: (A) Compared with the right hindlimb (R), the left hindlimb subjected to IR had increased protein expression of ChAT and CHT1 in tissue samples isolated from quadriceps femoris muscle within 16 h ( P <0.05, n = 6, respectively). (B) The heart excised after IR also showed increased expression of both ChAT and CHT1, and the levels were peaked at 12 h (0 h vs. 12 h: P <0.05, n = 6, respectively) in the heart.

Article Snippet: The following primary antibodies were used: a goat anti-ChAT polyclonal antibody (Millipore) at 1∶500 dilution; a rabbit anti-CHT1 polyclonal antibody (Antagene, Inc., Limonest, France) at 1∶500 dilution; and a rabbit anti-cleaved caspase-3 monoclonal antibody (Cell Signaling Technology, Inc.) at 1∶500 dilution.

Techniques: Expressing, Isolation