Journal: Nature Communications

Article Title: High-throughput screening identifies Aurora kinase B as a critical therapeutic target for Merkel cell carcinoma

doi: 10.1038/s41467-025-56504-7

Figure Lengend Snippet: A RNAi druggable genome screen demonstrating mean viability reduction following knockdown in MKL-2 (VP-MCC) vs. MCC26 (VN-MCC) cells (n = 3). All viabilities > 100 set to 100. Please refer to Supplementary Fig. : knockdown of both INCENP (n = 3) and survivin (n = 5) reduce VP-MCC, but not VN-MCC viability. B AURKB knockdown reduces MKL-2 and MCC26 viability relative to negative control (NC) (n = 3). Mean viability from 3 different siRNAs, error bars represent SEM. C Representative Western Blot demonstrating AURKB knockdown with siRNA (si), but not NC in MCC cell lines (n = 3). Red: β-actin loading control, green: AURKB. D Western Blot quantification demonstrating efficient AURKB siRNA knockdown relative to NC in WAGA (p = 0.001; n = 4), MKL-1 (p = 0.00003; n = 3), MCC13 (p = 0.00009; n = 3), and UISO (p = 0.045; n = 3) cell lines as measured by one-tailed two-sample t-test with unequal variance. AURKB intensity was normalized to β-actin intensity from identical lane, and means from at least 3 independent experiments are shown, with error bars representing SD. E AURKB knockdown significantly reduced viability of WAGA (p = 0.0003; n = 4), MKL-1 (p = 0.0105; n = 3), MCC13 (p = 0.0004; n = 3), and UISO (p = 0.0094; n = 3) cells relative to NC as measured by one-sample t-test. Data points are means from at least 3 independent experiments, with error bars representing SD. A and B, n = number of siRNAs per gene; C, D and E, n = number of experimental replicates.

Article Snippet: Paraffin-embedded MCC13 (VN-MCC) tissue sections were deparaffinized, rehydrated, and treated with Bloxall solution (SP-6000, Vector Laboratories, Burlingame, CA, USA) at room temperature to block endogenous peroxidase activity, followed by heat-mediate antigen retrieval using sodium citrate buffer (Bio SB, Santa Barbara, CA, USA).

Techniques: Knockdown, Negative Control, Western Blot, Control, One-tailed Test

Journal: Nature Communications

Article Title: High-throughput screening identifies Aurora kinase B as a critical therapeutic target for Merkel cell carcinoma

doi: 10.1038/s41467-025-56504-7

Figure Lengend Snippet: A Representative Fluorescent Western Blot of total histone 3 expression and pH3-Ser10. MCC cells were treated with DMSO, 3 nM, or 30 nM AZD2811 for 24 h (72 h for MCC26). Images were cropped to improve clarity (n = 3). B Quantification of pH3-Ser10 protein expression normalized to total histone 3 relative to DMSO control treatment from three independent experiments, with error bars representing SEM. 30 nM of AZD2811 significantly reduced pH3-Ser10 in WAGA (3 nM: p = 0.002, 30 nM: p < 0.0001; n = 6), MKL-2 (3 nM: p = 0.097, 30 nM: p = 0.00072; n = 3), MKL-1 (3 nM: p = 0.002, 30 nM: p < 0.0001; n = 3), MCC13 (3 nM: p = 0.043, 30 nM: p = 0.011; n = 5), and MCC26 (3 nM: p = 0.026, 30 nM: p = 0.031; n = 3) cells, as measured by two-tailed, two-sample, unpaired t-test. C Representative flow cytometry propidium iodide (PI) intensity histogram traces following treatment with 30 nM AZD2811 demonstrating a G2/M cell cycle arrest at 24 h in WAGA, MKL-2, MKL-1, and MCC13 and increased polyploidy ( > G2) at 24 and 72 h in MCC26 (n = 3). D AZD2811 24-h treatment (30 nM) significantly increased the proportion of cells in G2 in WAGA (p = 0.022; n = 3), MKL-1 (p = 0.0008; n = 3), MKL-2 (p = 0.016; n = 3), MCC13 (p = 0.0056; n = 3), but not MCC26 cells (p = 0.149; n = 3) as measured by two-sample, one-tailed unpaired t-test. AZD2811 treatment (30 nM) significantly increased the proportion of polyploid cells ( > G2) in MCC26 at 24 h (p = 0.0006; n = 3) and 72 h (p = 0.00759; n = 3). Values are means from at least 3 independent experiments with error bars representing SEM. n = number of experimental replicates.

Article Snippet: Paraffin-embedded MCC13 (VN-MCC) tissue sections were deparaffinized, rehydrated, and treated with Bloxall solution (SP-6000, Vector Laboratories, Burlingame, CA, USA) at room temperature to block endogenous peroxidase activity, followed by heat-mediate antigen retrieval using sodium citrate buffer (Bio SB, Santa Barbara, CA, USA).

Techniques: Western Blot, Expressing, Control, Two Tailed Test, Flow Cytometry, One-tailed Test

Journal: Nature Communications

Article Title: High-throughput screening identifies Aurora kinase B as a critical therapeutic target for Merkel cell carcinoma

doi: 10.1038/s41467-025-56504-7

Figure Lengend Snippet: A Schematic of xenograft tumor treatment protocol. Mice were subcutaneously injected with either MKL-1 or MCC13 cells (n = 10). Once the average tumor size reached 100 mm 3 animals were treated with intravenous AZD2811NP (25 mg/kg, twice weekly) or placebo nanoparticles for 4 weeks. B, C Rate of tumor growth, as calculated by linear regression, was significantly reduced by AZD2811NP treatment relative to placebo. P-values on the slope comparing vehicle and AZD2811NP were MKL-1 (p < 0.0001) and MCC13 (p < 0.0001). Data points represent average tumor volumes, with error bars representing SEM (n = 10). Please refer to Supplementary Fig. : AZD2811NP significantly slowed MKL-1 and MCC13 tumor growth relative to placebo during treatment period as measured by linear regression. Treatment with AZD2811NP did not reduce body weight in mice xenografted with MKL-1 (p = 0.71) or MCC13 (p = 0.9). D, E Kaplan Meier survival estimates showing that AZD2811NP extends survival of mice with MKL-1 (p < 0.0001) or MCC13 (p < 0.0001) xenograft tumors as calculated by Log-rank (Mantel-Cox) test in GraphPad Prism (n = 10). F, G Representative photographs of mice (MKL-1: day 28; MCC13: day 22) labeled with treatment. A , D , E , F , and G , n = number of animals; B and C , n = number of tumors.

Article Snippet: Paraffin-embedded MCC13 (VN-MCC) tissue sections were deparaffinized, rehydrated, and treated with Bloxall solution (SP-6000, Vector Laboratories, Burlingame, CA, USA) at room temperature to block endogenous peroxidase activity, followed by heat-mediate antigen retrieval using sodium citrate buffer (Bio SB, Santa Barbara, CA, USA).

Techniques: Injection, Labeling