g9a inhibitor unc0631  (MedChemExpress)

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: Lamin A/C maintains genome topology and regulates transcriptional programs essential for virus-driven B cell activation

doi: 10.64898/2026.01.12.699161

Figure Lengend Snippet: A) Overlap of deregulated genes (DEG) in RNA-Seq analysis between lamin A/C KO and G9a inhibitor treatment in WT cells. The overlap was statistically significant (p < 10 -16 ). B) Concordant regulation of the genes whose expression was altered by both lamin A/C KO and G9a inhibition. Data are represented using log 2 fold changes, and significantly regulated genes are depicted in red (upregulated) and blue (downregulated). The correlation coefficient (R = 0.73) indicates a strong positive correlation between the datasets. C) and D) H3K9me2 ChIP-Seq signal across the gene body of the 1,857 common targets between lamin A/C KO and G9a inhibition in WT cells. Increased deposition of H3K9me2 mark on the 927 targets that were downregulated in KO cells compared with WT (C), and no significant difference in H3K9me2 deposition on the 930 upregulated genes (D). Peaks at ± 3 kb from the transcription starting site (TSS) and the transcription end site (TES). E) and F) Ingenuity Pathway Analysis of the 1,857 common targets between lamin A/C KO and G9a inhibitor-treated WT cells, showing the top deregulated pathways (E) and upstream transcriptional regulators (F). Red indicates activation and blue inhibition. Data are shown as a function of the predicted z-score. G-I) Heat-maps of RNA-Seq data from WT, G9a inhibitor-treated WT, and lamin A/C KO cells showing genes in the PI3K, glycolysis, and c-myc signaling pathways whose expression was concordantly altered (false discovery rate [FDR] < 5%) by lamin A/C KO and G9a inhibition. Color code represents different fold change (FC) values, with upregulated genes shown in red and downregulated genes shown in blue.

Article Snippet: Treatment with the G9a inhibitor UNC0631 (Med Chem Express, HY-13808) was given 72 h before collection at a concentration of 1 μM.

Techniques: RNA Sequencing, Expressing, Inhibition, ChIP-sequencing, Activation Assay, Protein-Protein interactions

Journal: bioRxiv

Article Title: Lamin A/C maintains genome topology and regulates transcriptional programs essential for virus-driven B cell activation

doi: 10.64898/2026.01.12.699161

Figure Lengend Snippet: A) Drug screening assay on WT and lamin A/C KO cells treated with a library of PI3K inhibitors. Each compound was tested at 10, 1, 0.1, and 0.01 μM. Cell viability was assessed by measuring luminescence and converting the luminescence values to % toxicity. The graph shows the correlation between the log IC 50 in WT and KO cells for each compound. B) Acalisib dose-response curve in WT and lamin A/C KO cells. Toxicity was calculated using a luminescence assay, where the luminescence values were converted to % toxicity. Treatment with bortezomib (1 μM) was included as a positive control (100% toxicity); 0% toxicity corresponded to the RLU values of negative controls (0.1% DMSO). IC 50 values for acalisib in WT and lamin A/C KO cells are indicated. C) Cell viability (% live cells) assessed by counting the number of live cells interacting with the green dye calcein AM in WT and lamin A/C KO cells after treating for 72 hours with the PI3K inhibitor (PI3Ki) acalisib (0.1 µM) or control (Ctrl, 0.1% DMSO). Data come from 2 experiments with 4 biological replicates each (*, p ≤ 0.05; **, p ≤ 0.01). Sample acquisition and analysis were conducted using a fluorescence microscope. Fluorescence was quantified using the ImageJ software. D) Cell viability (% live cells) assessed by counting the number of live WT cells interacting with calcein AM in samples treated with control (Ctrl, 0.1% DMSO), a G9a inhibitor (G9ai, 1 µM), acalisib (PI3Ki, 0.01 µM), or their combination. Data come from 2 experiments with 4 biological replicates each (*, p ≤ 0.05; **, p ≤ 0.01) Sample acquisition and analysis were conducted using a fluorescence microscope. Fluorescence was quantified using the ImageJ software. E) Representative fluorescence microscope images of WT cells treated for 72 hours with control (Ctrl, 0.1% DMSO), G9a inhibitor (1 µM), acalisib (0.01 µM), or their combination and stained using calcein AM.

Article Snippet: Treatment with the G9a inhibitor UNC0631 (Med Chem Express, HY-13808) was given 72 h before collection at a concentration of 1 μM.

Techniques: Drug discovery, Luminescence Assay, Positive Control, Control, Fluorescence, Microscopy, Software, Staining