Journal: The Journal of Physiological Sciences : JPS

Article Title: Genetic inactivation of TRPM4 does not alter the temperature-dependent movement of mouse microglia

doi: 10.1016/j.jphyss.2026.100067

Figure Lengend Snippet: Generation of Trpm4 -knockout mouse line. A. Map of the mouse Trpm4 locus showing exons 15 and 16 targeted for deletion by sgRNAs. Top, WT allele. Bottom, KO allele. The sgRNAs were designed to target introns 14 and 16 flanking exons 15 and 16. Specific primers for amplifying the WT(Fw2/Rv) or KO (Fw1/Rv) allele were used. B. A representative result of genotyping PCR using a mixture of the Fw1, Fw2, and Rv primers. Two fragments were amplified from heterozygote mouse samples, each indicating the WT or KO fragment. The bottom fragment (695 bp) was amplified using the WT primer pair. The top fragment (928 bp) was amplified using the KO primer pair, indicating successful deletion of exons 15 and 16, which was confirmed by direct sequencing. (C and D). The expression patterns of Trpm4 gene in mouse brain (C) or primary microglia (D) using Gapdh as a control. In (C), “M4-plasmid” indicates an amplified fragment using mouse TRPM4 plasmid as a template. E. Western blot analysis of Trpm4 in mouse TRPM4-expressing HEK293T cells (M4-HEK) and mouse colon samples from WT or TRPM4KO mice. The expected molecular weight of Trpm4 is 134 kDa.

Article Snippet: After transferring the proteins to a nitrocellulose membrane, the membrane was blocked for 1 h at room temperature and then incubated with anti-TRPM4 antibody (1:300, ACC044, Alomone) overnight at 4 °C.

Techniques: Knock-Out, Amplification, Sequencing, Expressing, Control, Plasmid Preparation, Western Blot, Molecular Weight

Journal: The Journal of Physiological Sciences : JPS

Article Title: Genetic inactivation of TRPM4 does not alter the temperature-dependent movement of mouse microglia

doi: 10.1016/j.jphyss.2026.100067

Figure Lengend Snippet: Genetic elimination of Trpm4 does not alter the temperature-dependent microglia movement. A. Trajectories of primary microglia from a representative preparation of WT and TRPM4KO mice recorded for 2 h at 33 °C, 37 °C, and 40 °C. Paths are arranged to show origins at x (horizontal axis) = y (vertical axis) = 0. Each line indicates the trajectory of a single cell. n indicates the number of cells analyzed per preparation. B. The average distances of migrating microglia isolated from WT or TRPM4KO mice exposed to 33 °C (WT, n = 117; TRPM4KO, n = 151), 37 °C (WT, n = 235; TRPM4KO, n = 240), or 40 °C (WT, n = 150; TRPM4KO, n = 175) were measured. Open circles indicate the migration distance of each microglia over 2 h. Horizontal lines indicate means ± SEM. At 33 °C, 37 °C, and 40 °C, WT microglia moved 112.94 ± 6.1 μm, 175.28 ± 5.24 μm, and 204.31 ± 7.27 μm, respectively, whereas TRPM4KO microglia moved 113.46 ± 5.1 μm, 175.28 ± 4.13 μm, and 201.35 ± 5.61 μm, respectively. **P < 0.01 (two-way ANOVA followed by post hoc Bonferroni test for multiple comparisons).

Article Snippet: After transferring the proteins to a nitrocellulose membrane, the membrane was blocked for 1 h at room temperature and then incubated with anti-TRPM4 antibody (1:300, ACC044, Alomone) overnight at 4 °C.

Techniques: Single Cell, Isolation, Migration

Journal: Nature Communications

Article Title: Noncanonical calcium-independent TRPM4 activation governs intestinal fluid homeostasis

doi: 10.1038/s41467-025-68014-7

Figure Lengend Snippet: A Fluorescence images stained with Transient Receptor Potential Cation Channel Subfamily M Member 4 (TRPM4, green) and intestinal epithelial maker, Cytokeratin-18 (CK-18, orange) in colon sections derived from wild-type (WT) (upper panel) and Trpm4 -/- (lower panel) mice. Hoechst staining (blue) was used to visualize the nuclei. Scale bar = 50 µm. B Procedure to investigate the laxative effect of DAB in mice. Fecal water content ( C ) and Bristol score ( D ) of the stool collected from WT and Trpm4 -/- mice between 24 and 36 h after Veh or DAB exposure (n = 7 mice/group; four male and three female). Data are presented as Mean ± SEM. **, p < 0.0001, vs . WT-Veh; ## , p < 0.0001, vs . WT-DAB. Fecal water content ( E ) and Bristol score ( F ) of the stool collected from Trpm4 flox/flox and Trpm ΔIEC mice (n = 7 mice/group; four male and three female). **, p < 0.0001, vs . Trpm4 flox/flox -Veh; ## , p < 0.0001, vs . Trpm4 flox/flox -DAB. Two-way ANOVA followed by Bonferroni’s multiple comparisons test. Figure 1B was Created in BioRender. https://BioRender.com/8h3ia6w .

Article Snippet: Primary antibodies against CK-18 (#bsm-33101M, RRID: AB_3678929) and TRPM4 (#ACC-044, RRID: AB_2040250) were from Bioss Antibodies (Beijing, China) and Alomone Labs (Jerusalem, IL, USA), respectively.

Techniques: Fluorescence, Staining, Derivative Assay