ck666  (Tocris)

Journal: Cell reports

Article Title: The actin and microtubule network regulator WHAMM is identified as a key kidney disease risk gene

doi: 10.1016/j.celrep.2025.115462

Figure Lengend Snippet: (A) Percent of cytotoxicity of primary tubule cells (TECs) treated with PBS or cisplatin in the presence or absence of CK666 (CK) or latrunculin A (LatA). (B) Left: immunoblots of cleaved caspase-3 (C-CASP3), full-length gasdemrin D (F-GSDMD), cleaved GSDMD (N-GSDMD) protein, and GAPDH in TECs treated with PBS, cisplatin, or CK666 plus cisplatin. Right: quantification of immunoblots normalized to relative GAPDH level. (C) Left: confocal imaging of F-actin and ASC in Whamm +/+ and Whamm −/− TECs treated with cisplatin, and in Whamm +/+ TECs treated with CK666 plus cisplatin. Scale bars, 20 μm. Right: ImageJ analysis for the colocalized region (arrow): (upper) Whamm +/+, (middle) Whamm −/−, (lower) Whamm +/+ with CK666. (D) Quantification of ASC specks in TECs treated with cisplatin or CK666 plus cisplatin per region of interest (ROI). (E) WHAMM transcript levels in human renal proximal tubular epithelial cells (RPTECs) transduced with sgWHAMM or non-target (sgNST) virus. (F) Immunoblot of WHAMM in RPTECs after CRISPRi-mediated WHAMM silencing. (G) Quantification of WHAMM immunoblot normalized to GAPDH level. (H) F-Actin staining in RPTECs after CRISPRi-mediated silencing of WHAMM. Scale bar, 20 μm. Box indicates zoomed area. (I) ImageJ analysis of F-actin density and actin branch features. (J) WHAMM transcript levels in RPTECs following CRISPRi-mediated WHAMM silencing and treated with 40 μM of cisplatin for 24 h. (K) Percent of cytotoxicity in RPTECs after sg WHAMM or sgNST infection and treated with cisplatin for 24 h. (L) Transcript levels of BAX , CASPASE3 (CASP3) , CASP9 , and CASP1 , interleukin 1β ( IL-1B ), and IL-18 in RPTECs after sg WHAMM or sgNST infection and treated with cisplatin for 24 h. Data are presented mean ± SEM. p values were determined by one-way ANOVA or unpaired t test in GraphPad Prism 10 software. * p < 0.05, *** p < 0.001, **** p < 0.0001. See also .

Article Snippet: CK666 (#29038, Cayman) injections were continued for the rest of the two days of cisplatin-AKI and sacrificed on day 3 rd .

Techniques: Western Blot, Imaging, Transduction, Virus, Staining, Infection, Software

Journal: Cell reports

Article Title: The actin and microtubule network regulator WHAMM is identified as a key kidney disease risk gene

doi: 10.1016/j.celrep.2025.115462

Figure Lengend Snippet: (A) Experimental scheme of generating the CK666 (CK)-cisplatin model (CK-Cis). (B) Serum creatinine (sCr) and blood urea nitrogen (BUN) in the CK-Cis model ( n = 3–6 mice per group). (C) Transcript levels of Havcr1 , and Lcn2 in the CK-Cis model. (D) Whamm transcript level in the CK-Cis model. (E) H&E and PAS staining in kidney sections from the CK-Cis model. Scale bar, 50 μm. (F) The proposed mechanism for WHAMM in kidney disease. (G) Transcript levels of Nlrp3 , IL-1β , and IL-18 in the CK-Cis model or mice fed on adenine or control diet (CPD) (n = 4–5 mice per group). (H) Immunoblots of NLRP3, CASP1, cleaved CASP1 (P20), GSDMD-F, GSDMD-N, and GSDME in the CK-Cis model. (I) Immunoblots for cleaved GSDME and GAPDH in kidneys of control ( n = 3) or adenine-fed mice ( n = 6) injected with CK666 ( n = 5). (J) Quantification of NLRP3, CASP1, cleaved CASP1 (P20), GSDMD-F, GSDMD-N, and cleaved GSDME immunoblots in the CK-Cis model. Data are presented mean ± SEM. p values were determined by one-way ANOVA in GraphPad Prism 10 software. * p < 0.05, *** p < 0.001, **** p < 0.0001. See also .

Article Snippet: CK666 (#29038, Cayman) injections were continued for the rest of the two days of cisplatin-AKI and sacrificed on day 3 rd .

Techniques: Staining, Control, Western Blot, Injection, Software

Journal: bioRxiv

Article Title: Spatial distribution of cytoskeleton-mediated feedback controls cell polarization: a computational study

doi: 10.1101/2025.04.12.648264

Figure Lengend Snippet: (A-D) Differentiated neutrophils with F-actin-rich fronts (denoted by LifeAct-miRFP703) were imaged over time, and their shape and displacement were measured. Corresponding perturbations were simulated using Model 1 (local inhibition) and Model 2 (global inhibition), with the first 400 seconds showing STEN activity before effector recruitment (A–B) or drug addition (C–D), and the last 500 seconds showing STEN activity post-effector recruitment (A– B) or drug addition (C–D). The three colors in the kymograph represent Ras (Red), PKB (Blue), and PIP2 (Green). A. A 488 nm laser was switched on at frame 50 to globally recruit cytosolic CRY2PHR-mCherry-KRas4B G12V ΔCAAX to the cell periphery, increasing Ras activity and activating the STEN network. B. A 488 nm laser was switched on at frame 47 to globally recruit cytosolic CRY2PHR-mCherry-INP54P to the cell periphery, reducing PIP2 levels, which serves as the back molecule in STEN in both models. C. Differentiated neutrophils were pre-treated with 50 µ M CK666 to inhibit branched actin (denoted by LifeAct-miRFP703), turning off its positive feedback. D. Cells were pre-treated with 5 µ M Latrunculin B to completely remove actin polymerization (denoted by LifeAct-miRFP703), turning off both positive and negative feedback. Scale bar: 5 µ m.

Article Snippet: 10 μ M Latrunculin B (Sigma-Aldrich; Cat #428020) or 50 mM CK666 (EMD Millipore; Cat #182515) stock was prepared in dimethyl sulfoxide (DMSO, Sigma-Aldrich; Cat #D2650).

Techniques: Inhibition, Activity Assay

Journal: The Journal of Cell Biology

Article Title: Rectification of planar orientation angle switches behavior and replenishes contractile junctions

doi: 10.1083/jcb.202309069

Figure Lengend Snippet: Drug treatment reagents and concentrations

Article Snippet: CK666 (Cat#3950; TOCRIS) , Arp2/3 inhibitor , DMSO diluted 1:20 in H 2 O , 250 μm.

Techniques: Concentration Assay