Fig 1: Knocking down PAF1 affects radiosensitivity of CC by regulating IER5. a&b qRT-PCR assays of mRNA expression in radiation-exposed and unexposed Siha and Hela cells showing increased expression of IER5 and reduced PAF1 after radiation. β-actin served as expression controls. Data are expressed as the mean (S.D.) (n = 3). c&d Western blotting analysis results showing that based on radiation, knocking down PAF1 promoted the expression of IER5, while knocking down IER5 has no significant effect on PAF1. Fold change of intensity is mentioned in the western blotting bands. GAPDH served as expression controls. e&f FCM assay for analyzing the percentage of cells with active caspase-3 showing that PAF1 knockdown resulted in an increased percentage of cell with active caspase-3 and simultaneous PAF1 and IER5 knockdown resulted in little effect on the apoptosis proportion. N = 3 in each group. All data are expressed as the mean (S.D.): * P < 0.05 and ** P < 0.01
Fig 2: Effect of Met in combination with chemotherapy drugs on caspase activity. Percentage of caspase-1, −3, −4, −5, −6, −7, −8, and −9 activity (TF2-VAD-FMK) in cells treated with Met in combination with (A) chemotherapy drugs and (B) chemotherapy regimens. Percentage of caspase-3 activity (FITC-DEVD-FMK) in cells treated with Met in combination with (C) chemotherapy drugs and (D) chemotherapy regimens. Percentage of caspase-8 activity (FITC-IETD-FMK) in cells treated with Met in combination with (E) chemotherapy drugs and (F) chemotherapy regimens. Percentage of caspase-9 activity (FITC-LEHD-FMK) in cells treated with Met in combination with (G) chemotherapy drugs and (H) chemotherapy regimens. Data are presented as the mean ± SD from three independent experiments performed in triplicate. *P<0.05 vs. control; #P<0.05. Met, metformin; Epi, epirubicin; Cis, cisplatin; Dtx, docetaxel; 5FU, 5-fluorouracil; ECF, epirubicin + cisplatin + 5-fluorouracil; DCF, docetaxel + cisplatin + 5-fluorouracil; CF, cisplatin + 5-fluorouracil; ns, not significant.
Fig 3: The function of PSAT1 in xenograft models. (a) Combined endogenous (PSAT1 knockdown) and exogenous (serine‐deficient diet) serine reduction synergistically inhibited NSCLC tumor growth. (b) Mice receiving serine‐deficient diets (NC‐SL, shA‐SL) showed significant weight loss (n = 5, p < 0.05). (c) Tumor burden was lowest in the shA‐SL group (n = 5, p < 0.05). (d) IHC confirmed PSAT1 knockdown (shA groups), with Ki67 staining showing reduced proliferation and Caspase 3/TUNEL staining indicating increased apoptosis under serine deprivation.
Fig 4: CJE induces apoptosis and DNA damage in NK cells.PBMCs were exposed or not to 3 µg/mL of CJE. (A) Activated caspase-3 levels were measured in CD3 ⁻ CD56 ⁺ NK cells using the Caspase-3 Detection Kit and flow cytometry. (B) DNA damage was assessed by quantifying γH2AX expression in NK cells via flow cytometry. Data are presented as mean ± SEM. *p < 0.05; **p < 0.01; n = 4-5 independent experiments. Untreated = no exposure; DMSO = dimethyl sulfoxide; CJE = cannabis joint extract; MFI = median fluorescence intensity.
Supplier Page from Abcam for Cleaved Caspase-3 Staining Kit (FITC)