Journal: bioRxiv

Article Title: PREMATURE BIRTH AND CESAREAN SECTION AFFECT NEONATAL CD4 + T CELL GENE EXPRESSION AND CELLULAR FUNCTION

doi: 10.64898/2026.02.21.707199

Figure Lengend Snippet: (A) We assessed T cell activation by measuring the expression of CD69 and CD25 on the cell surface. Purified CD4⁺ T cells from term and preterm neonates were either left unstimulated or stimulated under the indicated conditions for 72 hours. The percentages of CD69⁺ and CD25⁺ T cells were determined by flow cytometry. (B) For transcription factor activation, we evaluate phosphorylation of c-Jun and p65 as markers of AP-1 and NF-κB activation, respectively. CBMCs were either left unstimulated or stimulated under the indicated conditions for 90 minutes (to measure c-Jun activation) or 60 minutes (for p65 activation). Mean fluorescence intensity (MFI) values for the indicated phosphoproteins were determined by flow cytometry. (C) MFI normalized values. The Wilcoxon test was used for paired samples, and the Mann–Whitney U test was used for unpaired samples. P values are shown above the error bars. * P < 0.05, ** P < 0.01. For the evaluation of CD69 and CD25 expression, the number of samples was as follows: vaginal delivery = 13, cesarean section = 9, and preterm birth = 3. For the evaluation of transcription factor activation, the sample sizes were: Natural birth = 7, Cesarean = 8, and Preterm = 5.

Article Snippet: We used anti–phospho c-Jun (Ser63, cat. GTX61170, GeneTex), secondary anti–rabbit FITC-conjugated antibody (GeneTex), and anti–phospho p65 (Ser529)–APC (Miltenyi Biotec).

Techniques: Activation Assay, Expressing, Purification, Flow Cytometry, Phospho-proteomics, Fluorescence, MANN-WHITNEY

Journal: iScience

Article Title: Evolutionary adaptations of TRPA1 thermosensitivity and skin thermoregulation in vertebrates

doi: 10.1016/j.isci.2025.113369

Figure Lengend Snippet: Identification and expression of trpv and trpa family members in Xenopus laevis melanophores (A) Molecular phylogenetic analysis by the maximum likelihood method of Trpv and Trpa family members (not to scale) of the predicted proteins from the genes found in the X. laevis genome. The name, gene number and chromosome localization are indicated. Brackets denote genes duplicated and maintained on the long (L) and short (S) chromosomes, with single copy genes indicated with an asterisk. (B and C) Representative RT-PCR analysis of mRNA expression in stage 43/44 whole embryos (B), and for isolated tails and MEX cells (C). The expressed mRNAs (L or S) identified after sequencing are indicated. Representative example of 3 independent samples; N = 3. (D–F) Immunohistochemistry for Trpv1 (D, F) or Trpa1 (E, F) and the Tyrosinase related protein 1 (Tyrp1) in consecutive sections (D, E), and in MEX cells (F). DAPI staining (blue) was used to visualize cell nuclei and facilitate the overlapping of adjacent sections in D and E. Boxed areas are shown enlarged (D′ or E′ and D″ or E″). Scale bar in D = 100 μm. Immunolabel indicates co-localization in melanophores (M) of the skin (S) of Tyrp1 (arrows) with the Trp channels. Note, adjacent sections were used as all antibodies were generated in rabbits. Note that Tyrp1 is localized to melanosomes while the Trp channels are in the plasma membrane. Scale bar in F = 10 μm.

Article Snippet: Anti human TRPA1 (rabbit polyclonal) , Biorbyt , Cat #: orb575403.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Isolation, Sequencing, Immunohistochemistry, Staining, Immunolabeling, Generated, Clinical Proteomics, Membrane

Journal: iScience

Article Title: Evolutionary adaptations of TRPA1 thermosensitivity and skin thermoregulation in vertebrates

doi: 10.1016/j.isci.2025.113369

Figure Lengend Snippet: Melanosome dispersion is affected by a Trpa1 agonist and antagonist (A and B) Tadpoles (Stage 43/44) or MEX cells were treated with agonists (A) or antagonists (B) against the indicated Trp channels. The agonists were added at 16°C and maintained at that temperature, while antagonists were added at 16°C, and the embryos were then switched to 32°C for 45 min. Data points ( n = 9 embryos or n = 6 MEX cells pictures) and a boxplot (25 th to 75 th percentile) are represented; A representative study from three ( N = 3) independent experiments is shown, with Capsaicin ( N = 2). Statistical significances for the different compounds are indicated against the control without the drug at the same temperature. ns; non significant. ∗; p < 0.05. ∗∗; p < 0.01.∗∗∗; p < 0.001. ∗∗∗∗; p < 0.0001.

Article Snippet: Anti human TRPA1 (rabbit polyclonal) , Biorbyt , Cat #: orb575403.

Techniques: Dispersion, Control

Journal: iScience

Article Title: Evolutionary adaptations of TRPA1 thermosensitivity and skin thermoregulation in vertebrates

doi: 10.1016/j.isci.2025.113369

Figure Lengend Snippet: Melanosome dispersion induced by temperature is mediated by Trpa1 (A) MEX cells transfected with a GFP-expression vector (control) or siRNA oligonucleotides, sense (S) or antisense (AS) against Trpa1, together with GFP in a 5:1 ratio. GFP-expressing cells had aggregated (orange arrow) or dispersed (white arrow) pigment. Representative images show the merge between GFP (pink), DAPI (yellow), and brightfield (BF). Scale bar = 10 μm. (B) Quantification of melanosome dispersion in GFP-expressing cells observed by shifting cells from 16°C to 32°C for 1 h after 48 h of transfection. Data are expressed as the percentage of GFP positive cells with dispersed melanosomes relative to the total number of GFP positive MEX cells ( N = 3; n = 20 pictures). Numbers in the bars indicate the counted cells. p value indicates a Fisher’s exact test comparing S and AS at 32°C.

Article Snippet: Anti human TRPA1 (rabbit polyclonal) , Biorbyt , Cat #: orb575403.

Techniques: Dispersion, Transfection, Expressing, Plasmid Preparation, Control

Journal: iScience

Article Title: Evolutionary adaptations of TRPA1 thermosensitivity and skin thermoregulation in vertebrates

doi: 10.1016/j.isci.2025.113369

Figure Lengend Snippet: Representative TRPA1 Structure and Phylogenetic Diagram of Related Taxa (A) Schematic of the secondary structure of human TRPA1: The 16 ankyrin repeat domains (ARD) at the amino terminus are numbered, followed by the linkers, the six transmembrane domains (S1 to S6), the TRP like domain, and the coiled coil domain at the carboxy terminus. (B) Related taxa (classes or superfamily) of vertebrates with similar TRPA1 “thermosensation” as determined by electrophysiological data obtained from the literature are color coded (heat, red; cold, blue; temperature insensitive, black; and unknown, dashed circles). Extant species of interest shown separated (see Materials and Methods) into 26 groups corresponding to the colored taxonomic clades indicated on the right. Evolutionary years (millions), dividing different eras, are approximated and are indicated on the left. Teleost-specific duplication [TSD] is indicated in yellow. Blue circles denote the age of the oldest preserved fossils with fur or feathers with insulation capacity, likely related to the advent of homeothermy (groups 10 to 26). Groups 1 to 9 are ectotherms. Groups 14 to 23 (square) are analyzed in and contain several specific examples of marine mammals, while groups 24 to 26 (square) are analyzed in and contain taxonomic groups with cold-activated TRPA1 (group 25, some rodents) and temperature-insensitive TRPA1 (group 24, Sciuromorpha and lagomorphs; group 26, primates).

Article Snippet: Anti human TRPA1 (rabbit polyclonal) , Biorbyt , Cat #: orb575403.

Techniques: Insulation

Journal: iScience

Article Title: Evolutionary adaptations of TRPA1 thermosensitivity and skin thermoregulation in vertebrates

doi: 10.1016/j.isci.2025.113369

Figure Lengend Snippet: Comparative phylogenetic analysis of TRPA1 domains for three marine mammals with terrestrial relatives Molecular phylogenetic analysis by the maximum-likelihood method based on the JTT matrix model was performed with the amino acid sequences of species shown in . The trees with the highest log-likelihood are shown to scale. Note the scale changes between the more recent evolutionary event containing bears (polar bear; brown bear; black bear and panda) and seals (Pinnipeds; groups 21–23) with respect to the analysis with manatees (Sirenia; groups 14 to 16) and whales (Cetacea; groups 17 to 20). The full length TRPA1 sequence, the region containing the sensor with the ankyrin repeat domain (ARD) 1 to 9, ARDs 10 to 16, and the transmembrane domains (S1 to S6) were used for the analysis. Numbers in the right (mean ± SD) indicate the phylogenetic distance relative to that for full-length TRPA1. Lack of standard deviation indicates only one species with TRPA1 data available. Evolutionary analyses were conducted in MEGA X.

Article Snippet: Anti human TRPA1 (rabbit polyclonal) , Biorbyt , Cat #: orb575403.

Techniques: Sequencing, Standard Deviation

Journal: iScience

Article Title: Evolutionary adaptations of TRPA1 thermosensitivity and skin thermoregulation in vertebrates

doi: 10.1016/j.isci.2025.113369

Figure Lengend Snippet: Phylogenetic divergence and alignment of the S5 Region of Glires and Primates (groups 23 to 26) as indicated in (A) Phylogenetic analysis (not to scale) from all major rodent clades modified from Blanga-Kanfi and co-workers based on six nuclear genes. (B) S5 region alignment containing the glycine residue (G878) present in several rodents (Group 25: Myomorphas, Hystricomorphas, and Castorimorphas) that is essential for cold sensitivity. The substitution of this residue with valine (V) in primates (Group 26) and lagomorphs (Group 24: rabbits, hares, and pikas) correlates with, where data are available, instances of thermal differences in TRPA1 sensitivity. Substituting human V878 with G878, however, does not restore cold sensitivity. Note that TRPA1 in the rodent squirrel (Sciuromorpha) is temperature insensitive and contains V at amino acid 878 instead of G. Demonstrated electrophysiological data for TRPA1 thermal sensitivity (cold, blue; insensitive, black) are indicated on the right.

Article Snippet: Anti human TRPA1 (rabbit polyclonal) , Biorbyt , Cat #: orb575403.

Techniques: Modification, Residue