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: Veterinary and Animal Science

Article Title: Aerococcus viridans, Staphylococcus haemolyticus and Corynebacterium bovis : sub-inhibitory exposure of lactic acid and cross-resistance to β-lactams antibiotics

doi: 10.1016/j.vas.2026.100620

Figure Lengend Snippet: Trend of LA MIC changes for S. haemolyticus during sub-inhibitory treatment.

Article Snippet: Fig 5 dummy alt text While S. haemolyticus displayed a 2-fold rise of cefoperazon for 2 field isolates (4137SH and 3934SH) plus the DSMZ reference astrain, there was also a 1 step decrease for 1 isolate (3958SH).

Techniques:

Journal: Veterinary and Animal Science

Article Title: Aerococcus viridans, Staphylococcus haemolyticus and Corynebacterium bovis : sub-inhibitory exposure of lactic acid and cross-resistance to β-lactams antibiotics

doi: 10.1016/j.vas.2026.100620

Figure Lengend Snippet: Correlation of LA and ꞵ-lactam antibiotic* tolerance between pre- and post-exposure in S. haemolyticus . Tolerance compared in measured MIC steps (+1). ▼ = 1 step decrease from the initial MIC. One MIC-step equals to a 2-fold change in antibiotic MICs and a 1.3-fold change in LA MICs. *Cefquinome, penicillin + novobiocin, and amoxicillin + clavulanic acid showed no changes at all.

Article Snippet: Fig 5 dummy alt text While S. haemolyticus displayed a 2-fold rise of cefoperazon for 2 field isolates (4137SH and 3934SH) plus the DSMZ reference astrain, there was also a 1 step decrease for 1 isolate (3958SH).

Techniques:

Journal: Journal of Neurochemistry

Article Title: Validation of TRPA1 and TRPV1 Antibodies for Expression Detection in Mammalian Cells and Tissues

doi: 10.1111/jnc.70444

Figure Lengend Snippet: Calcium imaging and immunocytochemistry showing TRPV1 expression in transfected HEK‐293 cells. (A) Fura2 calcium imaging in HEK‐293 cells transiently expressing rTRPV1‐EYFP fusion protein. Representative traces of calcium transients evoked by capsaicin in EYFP + cells. Green traces correspond to EYFP + cells and gray traces to EYFP − cells. Pie chart represents the percentage of EYFP + cells that responded or failded to respond to capsaicin. Data obtained from 88 cells from 3 coverslips. (B–F left) Confocal images of TRPV1‐EYFP transiently expressed in HEK‐293 cells. EYFP (green), TRPV1 antibody (magenta) and Hoechst (HO, blue). Merge images correspond to the overlap between TRPV1 and EYFP on the left and Hoechst and brightfield (BF) on the right. Antibody dilution of the representative images is indicated in the lower right corner. Scale bar: 20 μm. (B–F right) Box plots display the specificity ratio (SR) for each tested dilution of the corresponding antibody. Each dot represents a single EYFP + cell. Boxes indicate the interquartile range (25th to 75th percentiles) and whiskers extend to the 5th and 95th percentiles. The horizontal line inside each box marks the median and the black dot indicates the mean. (*** p < 0.001 Mann–Whitney test). (G) Bar histogram graph summarizes the SR median difference between the tested antibody dilution and their controls lacking primary antibody (secondary antibody only; rabbit, goat or guinea pig) as calculated with the Hodges‐Lehman estimator. Error bars represent the 95% confidence interval. For each antibody and dilution, a minimum of 100 cells were analyzed across 4 fields from 2 independent transfections. $ Santa Cruz V1 antibody was generated against a human TRPA1 peptide.

Article Snippet: Alomone , TRPA1 , Rabbit , Alomone Labs , Human TRPA1 747–760. PC , 69.2% , ACC‐037 RRID:AB_2040232.

Techniques: Imaging, Immunocytochemistry, Expressing, Transfection, MANN-WHITNEY, Generated

Journal: Journal of Neurochemistry

Article Title: Validation of TRPA1 and TRPV1 Antibodies for Expression Detection in Mammalian Cells and Tissues

doi: 10.1111/jnc.70444

Figure Lengend Snippet: Calcium imaging and immunocytochemistry of TRPA1 expression in transfected HEK‐ 293 cells. (A) Representative traces of Fura2 calcium transients evoked by 50 μM AITC. Green traces correspond to tGFP + cells and gray traces to tGFP − cells. The pie chart represents the proportion of tGFP + cells that responded or failed to respond to AITC. Data obtained from 115 cells from 3 coverslips. (B–H left) Confocal images of TRPA1‐tGFP transiently expressed in HEK‐ 293 cells incubated with the indicated antibody. tGFP (green), TRPA1 (magenta) and Hoechst staining (blue). Merge images correspond to the overlap between TRPA1 and tGFP (lower‐left panel) or Hoechst and brightfield (HO, BF, lower‐right). Antibody dilution of the representative images is indicated in the lower right‐hand corner. Scale bar: 20 μm. (B–H right) box plots display the specificity ratio (SR) for each tested dilution of the corresponding antibody. Each dot represents a single tGFP + cell. Boxes indicate the interquartile range (25th to 75th percentiles) and whiskers extend to the 5th and 95th percentiles. The horizontal line inside each box marks the median and the black dot indicates the mean. (** p < 0.01, *** p < 0.001 Mann–Whitney test). (I) Bar histogram graph summarizes the SR median difference between the tested antibody dilution and their controls lacking primary antibody (secondary antibody only; rabbit or mouse) as calculated with the Hodges‐Lehman estimator. Error bars represent the 95% confidence interval. For each antibody and dilution, a minimum of 319 cells were analyzed across 4 fields from 2 independent transfections.

Article Snippet: Alomone , TRPA1 , Rabbit , Alomone Labs , Human TRPA1 747–760. PC , 69.2% , ACC‐037 RRID:AB_2040232.

Techniques: Imaging, Immunocytochemistry, Expressing, Transfection, Incubation, Staining, MANN-WHITNEY

Journal: Journal of Neurochemistry

Article Title: Validation of TRPA1 and TRPV1 Antibodies for Expression Detection in Mammalian Cells and Tissues

doi: 10.1111/jnc.70444

Figure Lengend Snippet: Western blot analysis for TRPV1 and TRPA1 antibodies specificity. (A) Immunoblots for TRPV1 using Abcam (1:1000), Millipore V1 (1:1000), Santa Cruz V1 (1:100), Neuromics GT (1:200), and Neuromics GP (1:1000) antibodies. (−) lanes: Untransfected HEK‐293 cells, (+) lanes: HEK‐293 cells transfected with rTRPV1‐EYFP (B) Immunoblots for TRPA1 using Aviva (1:300), Novus (1:500), Millipore A1 (1:500), Alomone (1:200), Proteintech (1:500), Sigma WH (1:500), and Santa Cruz A1 (1:100). (−) lanes: Untransfected HEK‐293 cells, (+) lanes: HEK‐293 cells transfected with hTRPA1‐tGFFP. For each antibody, top row: Immunoblot with each TRPV1/TRPA1 antibody. Middle row: EYFP/tGFP immunoblotting of the same membrane. Bottom image: GAPDH loading control. Each blot was performed at least three times to rule out technical artifacts in cases where no anti‐TRPV1/TRPA1 signal was detected. For every repetition, the identical lysate sample was probed with all antibodies. Full uncropped blots are available in the Figure . $ Santa Cruz V1 antibody was generated against a human TRPA1 peptide.

Article Snippet: Alomone , TRPA1 , Rabbit , Alomone Labs , Human TRPA1 747–760. PC , 69.2% , ACC‐037 RRID:AB_2040232.

Techniques: Western Blot, Transfection, Membrane, Control, Generated

Journal: Journal of Neurochemistry

Article Title: Validation of TRPA1 and TRPV1 Antibodies for Expression Detection in Mammalian Cells and Tissues

doi: 10.1111/jnc.70444

Figure Lengend Snippet: Calcium imaging and Immunochemistry of endogenous TRPA1 in cultured DRG neurons from TRPA1‐Cre‐ChR2‐EYFP mice. (A) Representative traces of calcium transients evoked by AITC and capsaicin in DRG neurons. Traces corresponding to cells responding to 50 μM AITC are displayed in yellow and non‐responding cells are displayed in gray. (B) The pie chart represents the proportion of neurons responding or failing to respond to AITC. Data obtained from 424 cells from 5 coverslips. (C–F) ICC or (G–J) IHC confocal images of DRG sensory neurons incubated with the indicated antibody. TRPA1 antibody (magenta) and βIIITubulin (cyan), merge images display TRPA1 antibody signal + βIII‐Tubulin. Dilution of the corresponding antibody shown in each example is indicated in the lower right corner. Scale bar: 50 μm. Representative images were chosen from 4 pictures of 2 different animals.

Article Snippet: Alomone , TRPA1 , Rabbit , Alomone Labs , Human TRPA1 747–760. PC , 69.2% , ACC‐037 RRID:AB_2040232.

Techniques: Imaging, Cell Culture, Incubation