Fig 2: Etoposide increased LEP expression and promotes cellular senescence in C3H10T1/2 cells (A) The expression of LEP was measured using qPCR. (B and C) Cellular senescence was assessed by β-galactosidase staining. (D and E) qPCR was performed to measure the expression of aging markers p21 and p16. (F) The protein levels of p21 and p16 were evaluated by western blot

Fig 3: LEP knockdown inhibited etoposide-induced cellular senescence in C3H10T1/2 cells (A) The expression of LEP was measured using qPCR. (B and C) Cellular senescence was assessed by β-galactosidase staining. (D and E) qPCR was performed to measure the expression of aging markers p21 and p16. (F) The protein levels of p21 and p16 were evaluated by western blot

Fig 4: OGT promoted O-GlcNAcylation of LEP at S50 site. (A) The O-GlcNAcylation level and the protein levels of OGT and OGA were measured by western blot. (B and C) The protein levels of OGT, OGA and LEP and the O-GlcNAcylation levels of LEP in C3H10T1/2 cells with OGT or OGA overexpression were measured by western blot. (D and E) The potential O-GlcNAcylation sites in LEP was predicted using DictyOGlyc-1.1 database. (F) After mutating the potential O-GlcNAcylation sites in LEP, the protein and O-GlcNAcylation levels of LEP was measured by western blot. (G) After C3H10T1/2 cells were treated with 10 µM cycloheximide for 0, 8, 16, and 24 h, the stability of LEP protein was measured by western blot

Fig 5: Analysis of the correlation between Leptin/AMPK and breast cancer patients in the TCGA database. Note: A GeneMANIA database predicts the Leptin/AMPK signaling pathway; B GEPIA analyzes the correlation between Leptin/AMPK (where the horizontal axis represents log2(PRKAA1 TPM), the vertical axis represents log2(LEP TPM), P-value < 0.05); C box plot of AMPK expression results in sequencing data (black represents the control group, breast cancer patients after chemotherapy without fatigue; red represents the observation group, breast cancer patients after chemotherapy with fatigue)