unc3230  (Cayman Chemical)

Journal: bioRxiv

Article Title: Depletion of BRCA1 Potentiates Progestin-Induced Cytoskeletal Changes in an Ovarian Cancer Cell Model

doi: 10.64898/2026.01.02.697409

Figure Lengend Snippet: (A) Signaling schematic representing the regulation by PIP5K1C/Rho/ROCK/Rac signaling. (B-I) Average migration of NE-1 and KO-BRCA1 pools in transwell migration assay. Vehicle (ethanol), 10nM R5020, or 10nM R5020 + inhibitor combo treatment for 6-hour migration timepoint, which includes a prior 24-hour pre-treatment of either vehicle (ethanol), 10nM R5020, or 10nM R5020 + inhibitor treatment, respectively. 10% FBS used as chemoattractant. Representative bright-field images to the side of each graph taken at 100µm magnification. (B-C) PIP5K1C Inhibitor: 10µM UNC3230. Graph represents the mean ± SD, * p = 0.0248, ** p ≤ 0.0082, *** p = 0.0001, **** p < 0.0001. (D-E) Rho Inhibitor: 1µg/ml CT04 (Note: no inhibitor pre-treatment). Graph represents the mean ± SD, * p = 0.0285, ** p ≤ 0.0095, **** p < 0.0001. (F-G) ROCK Inhibitor: 10µM Y-27632. Graph represents the mean ± SD, * p ≤ 0.0384, ** p ≤ 0.0016, **** p < 0.0001. (H-1) Rac Inhibitor: 25µM EHT1864. Graph represents the mean ± SD, * p = 0.0120, *** p = 0.0002, **** p < 0.0001.

Article Snippet: Where applicable, cells were treated with the following reagents at the indicated concentrations: 10nM R5020 (Perkin Elmer, NLP004005MG), 10μM UNC3230 (Med Chem Express, HY-110150), 1μg/ml Rho Inhibitor I (Cytoskeleton, Inc, CT04), 10μM Y27632 dihydrochloride (Med Chem Express, HY-10583), and 25μM EHT1864 (Med Chem Express, HY-16659).

Techniques: Migration, Transwell Migration Assay

Journal: iScience

Article Title: LPLAT11/MBOAT7-driven phosphatidylinositol remodeling ensures radial glial cell integrity in developing neocortex

doi: 10.1016/j.isci.2025.114248

Figure Lengend Snippet: Mboat7 KO cortex exhibits reduced PI(4,5)P 2 levels, and pharmacological inhibition of PI(4,5)P 2 synthesis induces Golgi rounding (A) Measurement of total PI in the cortices in Mboat7 +/− and Mboat7 −/− mice at E11.5–E13.5 using LC-MS/MS-based method ( n = 3 embryos [E13.5, Mboat7 −/− ], n = 4 embryos [E11.5 and E12.5 Mboat7 +/− ], and n = 5 embryos [E12.5 Mboat7 −/− and E13.5 Mboat7 +/− ] from two independent litters). Peak areas are normalized by the area of the internal standard (25:0 PI). Ara and non-Ara indicate arachidonic acid-containing and non-arachidonic-acid-containing species, respectively. (B, C) Measurement of total PI4P (B) and PI(4,5)P 2 (C) in the cortices in Mboat7 +/− and Mboat7 −/− mice at E11.5–E13.5 using SFC-MS/MS-based method ( n = 3 embryos [E11.5, E12.5], n = 6 embryos [E13.5 Mboat7 −/− ], and n = 7 embryos [E13.5 Mboat7 +/− ] from two independent litters). Peak areas are normalized by the area of internal standard (37:4 PI4P or 37:4 PI(4,5)P 2 ). (D–F) Imaging MS analysis of PI (D), PIP (E), and PIP 2 (F) at E13.5 cortices of Mboat7 +/− and Mboat7 −/− mice. Signals were normalized by total ion current. (G) Immunofluorescence staining for PI(4,5)P 2 in the cortices of Mboat7 +/− and Mboat7 −/− mice at E13.5. The right panels show zoomed-in views of the area within the dotted frame. (H) Quantitative analysis of PI(4,5)P 2 positive dots per area within 100-μm-wide bins ( n = 5 embryos [ Mboat7 +/− ] and n = 4 embryos [ Mboat7 −/− ] from two independent litters). (I) Immunofluorescence staining for GM130 in cultured E12.5 cortical hemispheres treated with DMSO or 1 μM PIPKIγ inhibitor (UNC3230). The Golgi apparatus is rounded in the hemispheres treated with UNC3230. The right panels show zoomed-in views of the area within the dotted frame. (J) Measurement of the lengths of the GM130 + Golgi apparatus in the ventricular zone within 50-μm-wide bins ( n = 4 hemispheres [DMSO] and n = 4 hemispheres [UNC3230] from two independent litters and two independent experiments) in (I). (K–O) UNC3230 was administered into the ventricle of wild-type mice at E12.5. PBS (containing 0.1% DMSO) was administered as the control group. The E13.5 cortices were immunostained for PI(4,5)P 2 (K), GM130 (L), E-cadherin (M), Sox2 (N), and p-H3 (O). (P) Quantitative analysis of PI(4,5)P 2 -positive dots per area within 100-μm-wide bins ( n = 4 embryos [PBS] and n = 6 embryos [UNC3230] from two independent litters). (Q) Graph shows the lengths of the GM130 + Golgi apparatus in the ventricular zone within 50-μm-wide bins ( n = 703 cells from 3 embryos [PBS] and n = 670 cells from 3 embryos [UNC3230] from two independent litters). (R) Ratio of apical intensity against total intensity from VZ to MZ is shown for E-cadherin ( n = 7 embryos [PBS] and n = 6 embryos [UNC3230] from two independent litters). (S) Quantitative analysis of apical and dispersed RGCs positive for p-H3 (Sox2 + p-H3 + cells) per area within 200-μm-wide bins ( n = 6 embryos [PBS] and n = 8 embryos [UNC3230] from two independent litters). (T) Immunofluorescence staining for GM130 in cultured E12.5 cortical hemispheres from Mboat7 +/− and Mboat7 −/− mice. Rounding of the Golgi apparatus was observed in Mboat7 +/− hemispheres treated with 10 μM LPLAT11 inhibitor (Sevenin-1). (U) Measurement of the lengths of the GM130 + Golgi apparatus in the ventricular zone within 50-μm-wide bins ( n = 5 hemispheres [DMSO, Mboat7 +/− ], n = 4 hemispheres [Sevenin-1, Mboat7 +/− ], n = 5 hemispheres [DMSO, Mboat7 −/− ], and n = 4 hemispheres [Sevenin-1, Mboat7 −/− ] from two independent litters and two independent experiments). Data are shown as mean ± SEM; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001; unpaired two-tailed Student’s t test (C, H, P, R, S), unpaired two-tailed Welch’s t test (J, Q), and one-way ANOVA with Tukey’s post hoc test (U). The color of the asterisks corresponds to the color of the respective groups in the graph. Scale bars, 20 μm [K, L, T, and enlarged figures in (G, I)]; 500 μm (D–F); 100 μm (others). See also and .

Article Snippet: UNC3230 , TargetMol , Cat# T23498.

Techniques: Inhibition, Liquid Chromatography with Mass Spectroscopy, Tandem Mass Spectroscopy, Imaging, Immunofluorescence, Staining, Cell Culture, Control, Two Tailed Test

Journal: bioRxiv

Article Title: The tubulin poly-glutamylase complex, TPGC, is required for phosphatidyl inositol homeostasis and cilium assembly and maintenance

doi: 10.1101/2025.03.03.641315

Figure Lengend Snippet: A ) sgCTL control, TBC1D19 -/- , and TTLL1 -/- cells were serum starved for 24 hr, and immuno-stained with antibodies against acetylated tubulin and INPP5E. ≥ 80 cilia were counted per sample in three independent experiments. Error bars, S.D. **** p ≤ 0.0001. Representative images were shown for each cell line. Scale bar, 1µm. ns, not significant. B ) sgRNA control and TBC1D19 knockout cells were serum starved for 48 hr, re-fed with serum-containing medium, fixed after 30 min, 2 hr, or 3 hr, and immuno-stained with GT335 antibody. ≥ 70 cells were counted per sample in two independent experiments. Error bars, S.D. *p ≤ 0.05, **p ≤ 0.01. C ) Ciliary resorption rate (or percentage reduction in ciliation) after serum addition was calculated based on data in panel B. Error bars, S.D. *p ≤ 0.05. D ) sgRNA control and TTLL1 knockout cells were treated and stained as in panel B. N ≥ 100 cells were counted per sample in two independent experiments. Error bars, S.D. ns, not significant. E ) sgRNA control and TBC1D19 knockout cells were serum starved for 24 hr, simultaneously treated with DMSO, 1μM LY-294002 (LY), 1μM or 10 μM UNC3230 (UNC), or 1μM or 10 μM ISA-2011B (ISA), as indicated, and immuno-stained with GT335. ≥ 70 cells were counted per sample in two independent experiments. Error bars, S.D. *p ≤ 0.05, **p ≤ 0.01. ns, not significant.

Article Snippet: UNC3230 (T15597) and ISA-2011B (T23498) were purchased from TargetMol.

Techniques: Control, Staining, Knock-Out