Journal: Frontiers in Immunology

Article Title: ULK1 Inhibition as a Targeted Therapeutic Strategy for Psoriasis by Regulating Keratinocytes and Their Crosstalk With Neutrophils

doi: 10.3389/fimmu.2021.714274

Figure Lengend Snippet: Inhibition of ULK1 suppresses proliferation and promotes apoptosis of keratinocytes in vitro . (A) Immunoblot of HaCat keratinocytes 24 hours after cocultured with DMSO, 5 µM or 10 µM SBI-0206965 (SBI). (B) Cell cycle analysis of HaCat keratinocytes treated with DMSO or 10 µM SBI for 24 hours. The bar graph shows the percentage of cell population in each phase of cell cycle. (C) Apoptosis of HaCat keratinocytes 24 hours after serum deprivation in the presence of DMSO or 10 µM SBI. (D) mRNA expression of psoriasis-related inflammatory mediators in HaCat keratinocytes cocultured with DMSO or 10 µM SBI for 24 hours. (E) mRNA and protein expression of ULK1 in HaCat keratinocytes transfected with negative control-siRNA(NC-siRNA) and ULK1-siRNA. (F) Cell cycle analysis of HaCat keratinocytes 72 hours after transfection with NC-siRNA or ULK1-siRNA. (G) Apoptosis of transfected HaCat keratinocytes 24 hours after serum deprivation. (H) mRNA expression of psoriasis-related inflammatory mediators in HaCat keratinocytes transfected with NC-siRNA or ULK1-siRNA for 72 hours. Data are presented as mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: Primary keratinocytes (PCS-200-010, ATCC) were cultured in Dermal Cell Basall Medium (PCS-200-030, ATCC) supplemented with Keratinocyte Growth Kit (PCS-200-040, ATCC), 10U/mL of penicillin and 10μg/mL of streptomycin and 25ng/mL of amphotericin (03-033-1B, BI).

Techniques: Inhibition, In Vitro, Western Blot, Cell Cycle Assay, Expressing, Transfection, Negative Control

Journal: Frontiers in Immunology

Article Title: ULK1 Inhibition as a Targeted Therapeutic Strategy for Psoriasis by Regulating Keratinocytes and Their Crosstalk With Neutrophils

doi: 10.3389/fimmu.2021.714274

Figure Lengend Snippet: Inactivation of ULK1 by SBI suppressed the inflammation in keratinocytes stimulated by neutrophil. (A) mRNA expression of psoriasis-related inflammatory mediators by keratinocytes stimulated with neutrophils isolated from healthy donors (HC) or (B) psoriasis patients in the presence of 10 µM DMSO or SBI for 10 hours. Data are representative of three independent experiments. Data are presented as mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

Article Snippet: Primary keratinocytes (PCS-200-010, ATCC) were cultured in Dermal Cell Basall Medium (PCS-200-030, ATCC) supplemented with Keratinocyte Growth Kit (PCS-200-040, ATCC), 10U/mL of penicillin and 10μg/mL of streptomycin and 25ng/mL of amphotericin (03-033-1B, BI).

Techniques: Expressing, Isolation

Journal: Frontiers in Immunology

Article Title: ULK1 Inhibition as a Targeted Therapeutic Strategy for Psoriasis by Regulating Keratinocytes and Their Crosstalk With Neutrophils

doi: 10.3389/fimmu.2021.714274

Figure Lengend Snippet: Autophagy inhibitors fail to fully replicate the effect of ULK1 inhibition on keratinocyte. (A–C) Expression of p62 and LC3 I/II in HaCat keratinocytes 24 hours after incubation with SBI0206965 (SBI) (A) or chloroquine (B) or 3-methyladenine(3-MA) at indicated concentration (C) . (D) Cell cycle analysis and (E) apoptosis of keratinocytes after 24 hours with the treatment of 10 µM chloroquine or 5 mM 3-MA. (F) mRNA expression of psoriasis-related inflammatory mediators by keratinocytes cocultured with neutrophils from healthy donors in the presence of 5 mM 3-MA. Data are representative of three independent experiments. Data are presented as mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

Article Snippet: Primary keratinocytes (PCS-200-010, ATCC) were cultured in Dermal Cell Basall Medium (PCS-200-030, ATCC) supplemented with Keratinocyte Growth Kit (PCS-200-040, ATCC), 10U/mL of penicillin and 10μg/mL of streptomycin and 25ng/mL of amphotericin (03-033-1B, BI).

Techniques: Inhibition, Expressing, Incubation, Concentration Assay, Cell Cycle Assay