TISSUE CULTURE PLATE, 12 WELL
- Known as:
- TISSUE CULTURE PLATE, 12 WELL
- Catalog number:
- TCP20-12
- Product Quantity:
- 1
- Category:
- -
- Supplier:
- BioBasic
- Gene target:
- TISSUE CULTURE PLATE 12 WELL
Related genes to: TISSUE CULTURE PLATE, 12 WELL
- Gene:
- LRRC8A NIH gene
- Name:
- leucine rich repeat containing 8 VRAC subunit A
- Previous symbol:
- LRRC8
- Synonyms:
- KIAA1437, FLJ10337, SWELL1
- Chromosome:
- 9q34.11
- Locus Type:
- gene with protein product
- Date approved:
- 2003-09-26
- Date modifiied:
- 2019-04-23
Related products to: TISSUE CULTURE PLATE, 12 WELL
Related articles to: TISSUE CULTURE PLATE, 12 WELL
- Blood pressure (BP) is positively correlated with the activity of volume-regulated chloride channels (VRCCs) in vascular smooth muscle cells (VSMCs) during hypertension, and LRRC8A has recently been identified as the key molecular component of VRCCs. However, the role of LRRC8A in BP regulation remains unclear. Here, we generated SMC-specific LRRC8A knockout (CKO) mice and performed bulk RNA-sequencing, ChIP-sequencing, and coimmunoprecipitation coupled with mass spectrometry to elucidate the underlying mechanisms. We found that systolic BP was largely reduced in CKO mice under basal conditions as well as in those with angiotensin II- or DOCA-salt-induced hypertension. Aortas from CKO mice exhibited decreased vasoconstriction, whereas vasodilation and cardiac function remained unaffected. Mechanistically, LRRC8A deletion reduced STAT1 degradation and enhanced Calcrl transcription, thereby strengthening the inhibitory effect of Calcrl on myosin light chain phosphorylation and identifying LRRC8A deficiency as a negative regulator of VSMC contractility. Moreover, LRRC8A deficiency promoted the interaction between CSRP2 and PIAS1 and subsequently suppressed STAT1 degradation. Pharmacological inhibition of STAT1 with fludarabine abolished the protective effects of LRRC8A deficiency against hypertension and vasoconstriction. In summary, our findings reveal an essential role for LRRC8A in BP regulation and highlight a potential therapeutic strategy for hypertension. - Source: PubMed
Publication date: 2026/09/22
Lu Feng-TingLv Xiao-FeiLiang Zhu-JunTang WeiLai Wen-YiYang Gui-YongLi Yan-ShanMu Yan-JunTang Yu-BoXing Hui-KunYang Zhong-HanGuan Yong-YuanZhou Jia-GuoChen Jian-HuiGuo Kai-MinLiang Si-JiaMa Ming-Ming - DCPIB is the prototypical inhibitor of leucine-rich repeat containing protein 8 volume-regulated anion channels (VRACs), yet the structural basis and state dependence of inhibition remain unclear. Here, we used chimeric channels, targeted mutagenesis, and electrophysiology to define the determinants of DCPIB inhibition and elucidate how drug binding is coupled to channel gating. Effective inhibition required coordinated contributions from extracellular loop 1 (EL1) and transmembrane domain 2 (TM2), which together form a functional module governing access to and stabilization of DCPIB-bound states. Within TM2, a single hydrophobic residue strongly influenced inhibition, indicating that subtle differences in helix packing shape drug sensitivity. Using the homo-heptameric 8C-8A(IL125) channel as a defined model, we found that DCPIB acts exclusively from the extracellular side, inhibits cooperatively, and enhances voltage-dependent inactivation. Mutational analysis showed that DCPIB interacts permissively with R103 of the outer constriction site (OCS) but does not depend on it for inhibition, suggesting that the inhibitor intercalates between subunits within a hydrophobic cleft to achieve stable pore engagement. Charged substitutions within TM2 provided functional evidence that DCPIB penetrates beyond the OCS. Mutations at positions implicated in lipid interactions markedly altered DCPIB efficacy without affecting volume sensitivity, indicating that productive drug-channel interactions, rather than lipid gating itself, are required for inhibition. Finally, charge neutralization on the conserved N-terminal constriction enhanced DCPIB sensitivity, supporting long-range coupling within the pore. Together, these results establish DCPIB as a state-dependent VRAC inhibitor that stabilizes an inactivated channel conformation and define mechanistic principles for the rational design of more potent and selective VRAC inhibitors. - Source: PubMed
Publication date: 2026/09/22
Yamada ToshikiKarakas ErkanDenton Jerod S - Volume-regulated anion channels (VRACs), formed by heteromers of LRRC8 subunits, mediate the transport of anions and organic osmolytes in numerous physiological processes. Although their gating mechanism remains poorly understood, conformational rearrangements of the cytosolic leucine-rich repeat domains (LRRDs) have been implicated as a key step towards activation. Here, we identify the intracellular subdomain (ISD) of the obligatory LRRC8A as a key element coupling LRRD motion to pore opening. Substitution of leucine 402 with tryptophan (L402W) within a hydrophobic pocket of the ISD rendered LRRC8A homomers constitutively open, independent of osmotic stimulation or pharmacological modulation. Mutation of the nearby tryptophan 168 to leucine (W168L) also enhanced channel activity whereas the combination of both mutations (W168L-L402W) partly attenuated the hyperactive phenotype of L402W. FRET measurements revealed increased LRRD flexibility of L402W, which was reduced in W168L-L402W or by coexpression with an inhibiting sybody. The cryo-EM structure of L402W showed pronounced ISD rearrangements and outward tilting of the LRRDs, while the double mutant W168L-L402W contains features explaining its intermediate phenotype. Together, these data identify the ISD as a regulatory hub that transmits conformational changes from the LRRDs to the transmembrane gate of VRAC. - Source: PubMed
Publication date: 2026/09/17
Bertelli SaraWang LuKlüssendorf MalteGavazzo PaolaPusch MichaelDutzler RaimundStauber Tobias - Volume-regulated anion channels, which are activated in response to hypotonicity, are heteromers composed of LRRC8 family members. They consist of the obligatory subunit LRRC8A and at least another paralog that determines their substrate preference. Here we investigate heteromeric assemblies composed of LRRC8A and D subunits, the latter of which confer permeability to amino acids, osmolytes and anti-cancer drugs. Using patch-clamp electrophysiology, we show that LRRC8A/D channels are reversibly activated by cell swelling and that their response is modulated by sybodies targeting the A subunits, which either potentiate or repress channel activity. Structures of the LRRC8A/D channel in complex with the inhibitory sybody Sb1 define the channel stoichiometry of four A and two D subunits, with Sb1 stabilizing the A subunits in a similar closed channel conformation as found in homomeric LRRC8A assemblies. In LRRC8A/D heteromers, the two D subunits weaken the arrangement of A subunits and thus enhance the activation properties of the channel. This channel conformation is strongly perturbed upon binding of the potentiating sybody Sb4, which disrupts the packing of the cytoplasmic domains linking their mobility to pore opening. - Source: PubMed
Publication date: 2026/09/17
Lehmann Elena FStierli FabienneRutz SonjaDeneka DawidDutzler Raimund - Despite recent therapeutic advances, platinum-based chemotherapy still remains a cornerstone in the treatment of muscle invasive bladder cancer (MIBC). We investigated the association between leucine-rich-repeat-containing 8A (LRRC8A) expression, a protein with a potential mechanistic role in platinum-sensitivity, and clinical outcomes following neoadjuvant chemotherapy in MIBC. - Source: PubMed
Publication date: 2026/08/25
Rytz Michèle AlinaHussung SaskiaFrehner LorenzBeyer JörgScheidewind LailaRoth BeatWidmer CarmenBaaral AavashFrancica PaolaRubin Mark ARottenberg SvenRodriguez-Calero AntonioAkhoundova Dilara