Journal: Frontiers in Immunology

Article Title: Yes-associated protein regulates autophagy to restore skin barrier function in atopic dermatitis

doi: 10.3389/fimmu.2025.1681148

Figure Lengend Snippet: The effect of YAP expression on skin barrier in AD mice. (A) Trans epidermal water loss (TEWL) and stratum corneum hydration (SCH) of each group. (B) YAP protein expression from immunohistochemistry with the quantification results shown in the graphs (×400), bar length = 50 μm. n = 3. (C) Filaggrin (FLG) protein expression from immunohistochemistry with the quantification results shown in the graphs (×400), bar length = 50 μm. n = 3. (D) FLG, involucrin (IVL), and loricrin (LOR) protein expression from western blot with the quantification results shown in the graphs. (E) YAP, p-YAP, mTOR, and p-mTOR protein expression from western blot with the quantification results shown in the graphs. Ctrl, control mice; AD, atopic dermatitis model mice; BLK, mice injected with empty vector lentivirus; YAP, mice injected with YAP overexpression lentivirus; AD+YAP, atopic dermatitis model mice injected with YAP overexpression lentivirus; sh-NC, mice injected with shRNA-NC lentivirus; sh-YAP, mice injected with YAP shRNA lentivirus; AD+sh-YAP, atopic dermatitis model mice injected with YAP shRNA lentivirus; AD+sh-YAP+RPM, AD+sh-YAP group with topical 0.2% rapamycin ointment. * P < 0.05, *** P < 0.001 Ctrl v.s. AD; # P < 0.05, ## P < 0.01, ### P < 0.001 AD+YAP v.s. AD; + P < 0.05, +++ P < 0.001 AD+sh-YAP v.s. AD; ^^ P < 0.01, ^^^ P < 0.001 AD+sh-YAP+RPM v.s. AD+sh-YAP.

Article Snippet: The primary antibodies used were YAP (1:1000, Cell Signaling Technology, #14074), p-YAPS127 (1:1000, Cell Signaling Technology, #13008), FLG (1:1000, Gene Tex, GTX23137), IVL (1:400, bioss, bs-23062R), LOR (1:1000, novus, NBP1-33610), LC3A/B (1:1000, Cell Signaling Technology, #12741S), p62 (1:1000, Cell Signaling Technology, #39749S), mTOR (1:500, wanleibio, WL02477), p-mTOR (1:500, wanleibio, WL03694), β-actin (1:1000, wanleibio, WL01372), Beclin-1 (1:1000, wanleibio, WL02508), ATG5 (1:1000, wanleibio, WL02411), and ATG7 (1:500, wanleibio, WL02793).

Techniques: Expressing, Immunohistochemistry, Western Blot, Control, Injection, Plasmid Preparation, Over Expression, shRNA, Ointment

Journal: Frontiers in Immunology

Article Title: Yes-associated protein regulates autophagy to restore skin barrier function in atopic dermatitis

doi: 10.3389/fimmu.2025.1681148

Figure Lengend Snippet: The effect of YAP expression on skin barrier in AD like inflammatory cell model. (A) The schematic overview of atopic dermatitis (AD) like inflammatory cell model. (B) Flow cytometry analysis of cell apoptosis rate results. (C) Cell apoptosis from flow cytometry analysis. (D) Cell viability from MTT analysis. (E) FLG, IVL, and LOR protein expression from western blot with the quantification results presented in the graphs. (F) YAP, p-YAP, mTOR, and p-mTOR protein expression from western blot with the quantification results presented in the graphs. Ctrl, control cells; AD, atopic dermatitis-like inflammatory cells; BLK, cells transfected with empty vector; YAP, cells transfected with YAP overexpression plasmid; AD+YAP, atopic dermatitis model cells transfected YAP overexpression plasmid; si-NC, cells transfected with si-NC; si-YAP, cells transfected with si-YAP; AD+si-YAP, atopic dermatitis model cells transfected with si-YAP; AD+si-YAP+RPM, AD+si-YAP group with 50 nM rapamycin co-culture. * P < 0.05, ** P < 0.01, *** P < 0.001 Ctrl v.s. AD; ## P < 0.01, ### P < 0.001 AD+YAP v.s. AD; + P < 0.05, ++ P < 0.01, +++ P < 0.001 AD+si-YAP v.s. AD; ^ P < 0.05, ^^^ P < 0.001 AD+si-YAP+RPM v.s. AD+si-YAP.

Article Snippet: The primary antibodies used were YAP (1:1000, Cell Signaling Technology, #14074), p-YAPS127 (1:1000, Cell Signaling Technology, #13008), FLG (1:1000, Gene Tex, GTX23137), IVL (1:400, bioss, bs-23062R), LOR (1:1000, novus, NBP1-33610), LC3A/B (1:1000, Cell Signaling Technology, #12741S), p62 (1:1000, Cell Signaling Technology, #39749S), mTOR (1:500, wanleibio, WL02477), p-mTOR (1:500, wanleibio, WL03694), β-actin (1:1000, wanleibio, WL01372), Beclin-1 (1:1000, wanleibio, WL02508), ATG5 (1:1000, wanleibio, WL02411), and ATG7 (1:500, wanleibio, WL02793).

Techniques: Expressing, Flow Cytometry, Western Blot, Control, Transfection, Plasmid Preparation, Over Expression, Co-Culture Assay