Journal: Journal of Oncology

Article Title: Palbociclib Enhances Migration and Invasion of Cancer Cells via Senescence-Associated Secretory Phenotype-Related CCL5 in Non-Small-Cell Lung Cancer

doi: 10.1155/2022/2260625

Figure Lengend Snippet: Blocking SASP-related CCL5 inhibits the migration ability induced by Palbociclib. (a) The effect of antagonists of CCR5 TAK-779 on the migration ability of NSCLC cells treated with 2 μ M Palbociclib. Upper: Images of migrated cell with Palbociclib or TAK-779, bar: 100 μ m. Lower: The results were analyzed using unpaired t -test. (b) Diagram showing how Palbociclib contributes to the migration and invasion of NSCLC via SASP-related CCL5.

Article Snippet: Antagonists of CCR5, TAK-779, was purchased from MCE (CA, USA).

Techniques: Blocking Assay, Migration

Journal: Genes to Cells

Article Title: Treatment With Ruxolitinib and TAK ‐779 Enhances GII .17 Human Norovirus Replication and Enables Serial Passaging in Human Intestinal Enteroids

doi: 10.1111/gtc.70139

Figure Lengend Snippet: Treatment with ruxolitinib and TAK‐779 enhances GII.17 replication in HIEs. J2 monolayers were pretreated with ruxolitinib (5 μM) and/or TAK‐779 (30 μM) for 3 h and then inoculated with a stool filtrate containing GII.17 (1.6 × 10 5 GEs/well) in a differentiation medium supplemented with the corresponding compounds and 500 μM GCDCA for 1 h at 37°C. After washing, the cells were cultured with the specified compounds and GCDCA. The cells and culture supernatants were collected at 24, 48, and 96 hpi, and viral RNA was quantified by RT‐qPCR. Differentiation medium containing DMSO and 500 μM GCDCA served as the vehicle control. Data are presented as the mean ± SD ( n ≥ 5). Statistical significance was assessed using the Kruskal–Wallis test followed by Steel's multiple‐comparison test. Asterisks indicate statistically significant differences compared with DMSO‐treated controls at each time point (* p < 0.05). ns, not significant; hpi, h postinfection.

Article Snippet: Briefly, J2 monolayers were pretreated with either ruxolitinib (5 μM; T1829, TargetMol, Boston, MA, USA), TAK‐779 (30 μM; T7499, TargetMol), both, or DMSO (control) at 37°C under 5% CO 2 for 3 h, and then inoculated with HuNoV filtrates—1.6 × 10 5 GEs/well for GII.17, 1.1 × 10 7 GEs/well for GII.4, and 4.3 × 10 5 GEs/well for GII.3—in a differentiation medium supplemented with 500 μM sodium glycochenodeoxycholate (GCDCA; Sigma‐Aldrich) with DMSO or the indicated compounds at 37°C for 1 h. The monolayers were then washed twice with complete medium without growth factors (CMGF[−]).

Techniques: Cell Culture, Quantitative RT-PCR, Control, Comparison

Journal: Genes to Cells

Article Title: Treatment With Ruxolitinib and TAK ‐779 Enhances GII .17 Human Norovirus Replication and Enables Serial Passaging in Human Intestinal Enteroids

doi: 10.1111/gtc.70139

Figure Lengend Snippet: Effects of ruxolitinib and TAK‐779 on GII.3 and GII.4 HuNoV replication in HIEs. J2 monolayers pretreated with ruxolitinib (5 μM) and/or TAK‐779 (30 μM) were infected with stool filtrate containing GII.3 (4.3 × 10 5 GEs/well) (a) or GII.4 (1.1 × 10 7 GEs/well) (b), as shown in Figure . At 48 hpi, the cells and culture supernatants were collected, and viral RNA was quantified by RT‐qPCR. Data are presented as the mean ± SD ( n ≥ 9). Statistical significance was assessed using the Kruskal–Wallis test followed by Steel's multiple‐comparison test. Asterisks indicate statistically significant differences compared with DMSO‐treated controls (** p < 0.01, **** p < 0.0001). hpi, h postinfection.

Article Snippet: Briefly, J2 monolayers were pretreated with either ruxolitinib (5 μM; T1829, TargetMol, Boston, MA, USA), TAK‐779 (30 μM; T7499, TargetMol), both, or DMSO (control) at 37°C under 5% CO 2 for 3 h, and then inoculated with HuNoV filtrates—1.6 × 10 5 GEs/well for GII.17, 1.1 × 10 7 GEs/well for GII.4, and 4.3 × 10 5 GEs/well for GII.3—in a differentiation medium supplemented with 500 μM sodium glycochenodeoxycholate (GCDCA; Sigma‐Aldrich) with DMSO or the indicated compounds at 37°C for 1 h. The monolayers were then washed twice with complete medium without growth factors (CMGF[−]).

Techniques: Infection, Quantitative RT-PCR, Comparison

Journal: Genes to Cells

Article Title: Treatment With Ruxolitinib and TAK ‐779 Enhances GII .17 Human Norovirus Replication and Enables Serial Passaging in Human Intestinal Enteroids

doi: 10.1111/gtc.70139

Figure Lengend Snippet: Treatment with ruxolitinib and TAK‐779 enables serial passaging of GII.17, and passaged viruses exhibit higher viral RNA copies than stool‐derived viruses in HIEs. (a) J2 monolayers pretreated with ruxolitinib (5 μM) and TAK‐779 (30 μM) were infected with stool filtrate containing GII.17 (1.6 × 10 5 GEs/well) in a differentiation medium supplemented with the specified compounds and GCDCA for 3 h at 37°C. At 96 hpi, the cells were harvested, sonicated, and centrifuged. A portion of the clarified supernatants was used for RT‐qPCR quantification of viral RNA. For serial passaging, the virus was used to inoculate newly prepared J2 monolayers under the same conditions for up to five passages. Data are presented as mean ± SD ( n ≥ 5). (b) J2 monolayers were inoculated with 4.6 × 10 4 GEs per well of stool‐derived GII.17 or P5 virus stock in differentiation medium supplemented with GCDCA for 1 h at 37°C. Following infection, cells and culture supernatants were collected at 48 hpi, and viral RNA was quantified by RT‐qPCR. Data are presented as mean ± SD ( n = 4). Statistical significance was assessed using the Welch t ‐test. Asterisks indicate statistically significant differences compared with stool‐derived infection at 48 hpi (* p < 0.05). hpi, h postinfection.

Article Snippet: Briefly, J2 monolayers were pretreated with either ruxolitinib (5 μM; T1829, TargetMol, Boston, MA, USA), TAK‐779 (30 μM; T7499, TargetMol), both, or DMSO (control) at 37°C under 5% CO 2 for 3 h, and then inoculated with HuNoV filtrates—1.6 × 10 5 GEs/well for GII.17, 1.1 × 10 7 GEs/well for GII.4, and 4.3 × 10 5 GEs/well for GII.3—in a differentiation medium supplemented with 500 μM sodium glycochenodeoxycholate (GCDCA; Sigma‐Aldrich) with DMSO or the indicated compounds at 37°C for 1 h. The monolayers were then washed twice with complete medium without growth factors (CMGF[−]).

Techniques: Passaging, Derivative Assay, Infection, Sonication, Quantitative RT-PCR, Virus

Journal: Genes to Cells

Article Title: Treatment With Ruxolitinib and TAK ‐779 Enhances GII .17 Human Norovirus Replication and Enables Serial Passaging in Human Intestinal Enteroids

doi: 10.1111/gtc.70139

Figure Lengend Snippet: Detection of GII.17 VP1 in HIEs by immunofluorescence. J2 monolayers pretreated with DMSO (a) or ruxolitinib (5 μM) and TAK‐779 (30 μM) (b) were infected with mock inoculum (left panels), stool filtrate containing GII.17 (middle panels), or P4 virus stock (right panels) in the presence of DMSO or specified compounds and GCDCA. At 96 hpi, the cells were fixed with 4% paraformaldehyde and permeabilized with 1% Triton X‐100. GII.17‐positive cells were detected using GII.17‐specific antiserum (green) and stained with DAPI (blue). Scale bars: 100 μm. DAPI, 4′,6‐diamidino‐2‐phenylindole; VP1, viral protein 1.

Article Snippet: Briefly, J2 monolayers were pretreated with either ruxolitinib (5 μM; T1829, TargetMol, Boston, MA, USA), TAK‐779 (30 μM; T7499, TargetMol), both, or DMSO (control) at 37°C under 5% CO 2 for 3 h, and then inoculated with HuNoV filtrates—1.6 × 10 5 GEs/well for GII.17, 1.1 × 10 7 GEs/well for GII.4, and 4.3 × 10 5 GEs/well for GII.3—in a differentiation medium supplemented with 500 μM sodium glycochenodeoxycholate (GCDCA; Sigma‐Aldrich) with DMSO or the indicated compounds at 37°C for 1 h. The monolayers were then washed twice with complete medium without growth factors (CMGF[−]).

Techniques: Immunofluorescence, Infection, Virus, Staining