(2r,3s,4r,5r,8r,10r,11r,12s,13s,14r)-11-{[(2s,3r,4s,6r)-4-(dimethylamino)-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(2r,4r,5s,6s)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-6-(2h3)methyl-3,5,8,10,12,14-hexamethyl-1-oxa-6-azacyclopentadecan-15-one  (Enamine Ltd)

Journal: bioRxiv

Article Title: Epilepsy-associated potassium channel KCNT1 is required for multiciliated cell development in Xenopus

doi: 10.64898/2026.03.31.710877

Figure Lengend Snippet: (A) Patient medical record data representing 46 patients with variants in KCNT1 reveal prevalence of clinical symptoms affecting many organ systems. (B) Within the respiratory issues, 78% of patients were dependent on assisted breathing, 48% experienced respiratory failure, 46% had excess fluid in their lungs, 41% experienced respiratory distress, 40% experienced hypoxia, and 24% had structural defects. (C) Within the cardiac issues, 40% of patients had abnormal ECG, 33% had heart murmurs, 28% had congenital structural heart defects, 13% had systemic-pulmonary collaterals, and 7% had arrhythmias. (D) Within the musculoskeletal defects, 59% of patients had hypotonia, 54% had hip deformities, 43% had joint contractures, 35% had spine deformities, and 33% had compromised bone integrity. (E) Within the kidney issues, 43% of patients had urinary dysfunction, 17% had recurrent UTIs, 15% had renal stones, 13% had urinary obstruction, and 4% had structural defects. (F) Within the dysmorphic features, 22% of patients had facial dysmorphic features, 20% had plagiocephaly, 11% had foot deformities, and 4% had leg length inequality. See also Table S1.

Article Snippet: KCNT1 antagonist (VU0606170, ProbeChem PC-73240), additional KCNT1 antagonist (Compound 31, Enamine EN300-27781598), Piezo1 antagonist (GsMTx4, MedChemExpress HY-P1410), and Piezo1 agonist (Yoda1, Sigma SML1558) were resuspended in DMSO at 10 mM, 10 mM, 5 mM, and 10 mM, respectively.

Techniques:

Journal: bioRxiv

Article Title: Epilepsy-associated potassium channel KCNT1 is required for multiciliated cell development in Xenopus

doi: 10.64898/2026.03.31.710877

Figure Lengend Snippet: (A) kcnt1 is expressed in ciliated embryonic tissues including the neural tube, epidermis, heart and pronephros in NF stages 20, 30, and 35 in X. tropicalis embryos. (B) kcnt1 (yellow, HCR probe) is broadly expressed on the ciliated epidermis, labeled with cell boundary protein E-cadherin (blue, antibody staining), cilia protein acetylated alpha-tubulin (green, antibody staining), and MCC marker foxj1 (magenta, HCR probe), in developing X. tropicalis embryos in NF stage 30. (C) Volcano plot of bulk RNA sequencing of embryos depleted of kcnt1 with translation-blocking morpholino in X. tropicalis with select significant genes, sequenced at stage 33, compared to control morpholino-injected animals. (D) Top 10 biological processes organized by adjusted p value followed by fold enrichment using GO term analysis of differentially expressed genes in C. See also Table S2, S3.

Article Snippet: KCNT1 antagonist (VU0606170, ProbeChem PC-73240), additional KCNT1 antagonist (Compound 31, Enamine EN300-27781598), Piezo1 antagonist (GsMTx4, MedChemExpress HY-P1410), and Piezo1 agonist (Yoda1, Sigma SML1558) were resuspended in DMSO at 10 mM, 10 mM, 5 mM, and 10 mM, respectively.

Techniques: Labeling, Staining, Marker, RNA Sequencing, Blocking Assay, Control, Injection

Journal: bioRxiv

Article Title: Epilepsy-associated potassium channel KCNT1 is required for multiciliated cell development in Xenopus

doi: 10.64898/2026.03.31.710877

Figure Lengend Snippet: (A) Depletion of kcnt1 by a translation-blocking morpholino (MO) reduces motile cilia compared to control morpholino-injected NF stage 28 X. tropicalis stained for acetylated-α-tubulin (cilia, magenta) and phalloidin (actin, grey), and injected with tracer H2B-GFP (nucleus, green). Scale bars = 200 µm. (B) Quantification of data shown in A as a percentage of embryos with no cilia phenotype vs. mild, moderate or severe cilia phenotype. See for representative images of cilia phenotype classifications. Embryo images were randomized and phenotyped blinded to condition. Chi-Square test was used to calculate significance using raw counts of individual embryos. **** = p < 0.0001. (C) Epidermis of embryos from A. Scale bars = 10 µm. (D) X. laevis embryonic treatment with KCNT1 antagonist VU0606170 reduces motile cilia compared to DMSO vehicle treated from NF stage 7 to NF stage 38 X. laevis . Embryos were stained with acetylated-α-tubulin (cilia, magenta) and phalloidin (actin, grey). Scale bars = 500 µm. (E) Quantification of data shown in D as a percentage of embryos with no cilia phenotype vs. mild, moderate or severe cilia phenotype. Chi-Square test was used to calculate significance using raw counts of individual embryos. **** = p < 0.0001. (F) Epidermis of embryos from D. Scale bars = 20 µm followed by 10 µm. See also for treatment with an additional KCNT1 inhibitor, Compound 31 , which also phenocopies these results.

Article Snippet: KCNT1 antagonist (VU0606170, ProbeChem PC-73240), additional KCNT1 antagonist (Compound 31, Enamine EN300-27781598), Piezo1 antagonist (GsMTx4, MedChemExpress HY-P1410), and Piezo1 agonist (Yoda1, Sigma SML1558) were resuspended in DMSO at 10 mM, 10 mM, 5 mM, and 10 mM, respectively.

Techniques: Blocking Assay, Control, Injection, Staining

Journal: bioRxiv

Article Title: Epilepsy-associated potassium channel KCNT1 is required for multiciliated cell development in Xenopus

doi: 10.64898/2026.03.31.710877

Figure Lengend Snippet: (A) Inhibition of KCNT1 with KCNT1 antagonist, Compound 31 (50 or 75 µM), reduces motile cilia compared to DMSO vehicle treated embryos, treated from NF stage 7 to NF stage 38 in X. laevis , stained with acetylated-α- tubulin (cilia, magenta) and phalloidin (actin, grey). Embryo images were randomized and scored blinded to condition. Scale bars = 500 µm. (B) Quantification of data shown in A as a percentage of embryos with no cilia phenotype vs. mild, moderate or severe cilia phenotype. Chi-Square test was used to calculate significance using raw counts of individual embryos. **** = p < 0.0001, *** = 0.0009. (C) Epidermis of embryos from A. Scale bars = 20 µm followed by 10 µm.

Article Snippet: KCNT1 antagonist (VU0606170, ProbeChem PC-73240), additional KCNT1 antagonist (Compound 31, Enamine EN300-27781598), Piezo1 antagonist (GsMTx4, MedChemExpress HY-P1410), and Piezo1 agonist (Yoda1, Sigma SML1558) were resuspended in DMSO at 10 mM, 10 mM, 5 mM, and 10 mM, respectively.

Techniques: Inhibition, Staining

Journal: bioRxiv

Article Title: Epilepsy-associated potassium channel KCNT1 is required for multiciliated cell development in Xenopus

doi: 10.64898/2026.03.31.710877

Figure Lengend Snippet: (A) Inhibition of KCNT1 and Piezo signaling with KCNT1 antagonist, VU0606170 (20 µM), and Piezo signaling antagonist, GsMTx4 (5 µM), further exacerbates loss of motile cilia compared to KCNT1 antagonist (20 µM) alone. X. laevis embryos were treated from NF stage 6 to 38 and stained with acetylated-α-tubulin (cilia, magenta) and phalloidin (actin, grey). Embryo images were randomized and scored blinded to condition. Scale bars = 500 µm. (B) Quantification of data shown in A as a percentage of embryos with no cilia phenotype vs. mild, moderate or severe cilia phenotype. Fisher’s exact test was used to calculate significance using raw counts of individual embryos. **** = p < 0.0001, ns = no statistically significant difference. (C) Partial rescue of loss of cilia phenotype from KCNT1 inhibition (20 µM VU0606170) by co-treatment with a Piezo1 agonist, Yoda1 (110 µM), compared to KCNT1 antagonist alone (20 µM). Scale bars = 500 µm. (D) Quantification of data shown in C as a percentage of embryos with no cilia phenotype vs. mild, moderate or severe cilia phenotype. Fisher’s exact test was used to calculate significance using raw counts of individual embryos. **** = p < 0.0001, *** = 0.0009, ns = no statistically significant difference.

Article Snippet: KCNT1 antagonist (VU0606170, ProbeChem PC-73240), additional KCNT1 antagonist (Compound 31, Enamine EN300-27781598), Piezo1 antagonist (GsMTx4, MedChemExpress HY-P1410), and Piezo1 agonist (Yoda1, Sigma SML1558) were resuspended in DMSO at 10 mM, 10 mM, 5 mM, and 10 mM, respectively.

Techniques: Inhibition, Staining

Journal: bioRxiv

Article Title: Epilepsy-associated potassium channel KCNT1 is required for multiciliated cell development in Xenopus

doi: 10.64898/2026.03.31.710877

Figure Lengend Snippet: (A) Inhibition of KCNT1 and Piezo signaling with KCNT1 antagonist, VU0606170 (20 µM), and Piezo signaling antagonist, GsMTx4 (5 µM), further exacerbates loss of motile cilia compared to KCNT1 antagonist (20 µM) alone. X. laevis embryos were treated from NF stage 6 to 38 and stained with acetylated-α-tubulin (cilia, magenta) and phalloidin (actin, grey). Embryo images were randomized and scored blinded to condition. Scale bars = 500 µm. (B) Quantification of data shown in A as a percentage of embryos with no cilia phenotype vs. mild, moderate or severe cilia phenotype. Fisher’s exact test was used to calculate significance using raw counts of individual embryos. **** = p < 0.0001, ns = no statistically significant difference. (C) Partial rescue of loss of cilia phenotype from KCNT1 inhibition (20 µM VU0606170) by co-treatment with a Piezo1 agonist, Yoda1 (110 µM), compared to KCNT1 antagonist alone (20 µM). Scale bars = 500 µm. (D) Quantification of data shown in C as a percentage of embryos with no cilia phenotype vs. mild, moderate or severe cilia phenotype. Fisher’s exact test was used to calculate significance using raw counts of individual embryos. **** = p < 0.0001, *** = 0.0009, ns = no statistically significant difference.

Article Snippet: KCNT1 antagonist (VU0606170, ProbeChem PC-73240), additional KCNT1 antagonist (Compound 31, Enamine EN300-27781598), Piezo1 antagonist (GsMTx4, MedChemExpress HY-P1410), and Piezo1 agonist (Yoda1, Sigma SML1558) were resuspended in DMSO at 10 mM, 10 mM, 5 mM, and 10 mM, respectively.

Techniques: Inhibition, Staining