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HCT116 parental cells and clones differing for the rs1053639 homozygous genotypes were treated with either Nutlin 10 µM or vehicle control for 16 hours. A) Cell lysates were analyzed by immunoblotting for the induction of p53 and DDIT4 protein levels and the phosphorylation state and levels of the eIF4EBP1 mTORC1 target. β-Tubulin and GAPDH were used as references. B, C) Densitometry analysis of p53 and DDIT4 protein levels shows significant induction upon Nutlin treatment. D) Densitometry analysis of <t>phosphorylated</t> eIF4EBP1 blots reveals that significant inhibition of the phosphorylation of eIF4EBP1 is apparent only in the rs1053639 TT homozygous clones. ***p-value <0.005. **p-value <0.001. *p-value <0.05. ns, not significant. unpaired t-test.
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<t>OXPHOS</t> protein expression. Densitometric analyses of subunits of complexes I (CI-NDUFB8, 20 kDa), II (CII-SDHB, 30 kDa), III (CIII-UQCRC2, 48 kDa) and V (CV-ATP5A, 55 kDa) of the OXPHOS system normalised to total proteins. A representative Western blot image of OXPHOS and total proteins is shown on the right side of the figure.
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<t>OXPHOS</t> protein expression. Densitometric analyses of subunits of complexes I (CI-NDUFB8, 20 kDa), II (CII-SDHB, 30 kDa), III (CIII-UQCRC2, 48 kDa) and V (CV-ATP5A, 55 kDa) of the OXPHOS system normalised to total proteins. A representative Western blot image of OXPHOS and total proteins is shown on the right side of the figure.
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HCT116 parental cells and clones differing for the rs1053639 homozygous genotypes were treated with either Nutlin 10 µM or vehicle control for 16 hours. A) Cell lysates were analyzed by immunoblotting for the induction of p53 and DDIT4 protein levels and the phosphorylation state and levels of the eIF4EBP1 mTORC1 target. β-Tubulin and GAPDH were used as references. B, C) Densitometry analysis of p53 and DDIT4 protein levels shows significant induction upon Nutlin treatment. D) Densitometry analysis of phosphorylated eIF4EBP1 blots reveals that significant inhibition of the phosphorylation of eIF4EBP1 is apparent only in the rs1053639 TT homozygous clones. ***p-value <0.005. **p-value <0.001. *p-value <0.05. ns, not significant. unpaired t-test.

Journal: bioRxiv

Article Title: A TranSNP in the DDIT4 mRNA can impact its translation efficiency and modulate p53-dependent responses in cancer cells

doi: 10.1101/2025.04.02.646512

Figure Lengend Snippet: HCT116 parental cells and clones differing for the rs1053639 homozygous genotypes were treated with either Nutlin 10 µM or vehicle control for 16 hours. A) Cell lysates were analyzed by immunoblotting for the induction of p53 and DDIT4 protein levels and the phosphorylation state and levels of the eIF4EBP1 mTORC1 target. β-Tubulin and GAPDH were used as references. B, C) Densitometry analysis of p53 and DDIT4 protein levels shows significant induction upon Nutlin treatment. D) Densitometry analysis of phosphorylated eIF4EBP1 blots reveals that significant inhibition of the phosphorylation of eIF4EBP1 is apparent only in the rs1053639 TT homozygous clones. ***p-value <0.005. **p-value <0.001. *p-value <0.05. ns, not significant. unpaired t-test.

Article Snippet: Lysates prepared for analysis of phosphorylated proteins were supplemented by a phosphatase inhibitor cocktail (Phosphatase Inhibitor Cocktail I, MedChemExpress).

Techniques: Clone Assay, Control, Western Blot, Inhibition

OXPHOS protein expression. Densitometric analyses of subunits of complexes I (CI-NDUFB8, 20 kDa), II (CII-SDHB, 30 kDa), III (CIII-UQCRC2, 48 kDa) and V (CV-ATP5A, 55 kDa) of the OXPHOS system normalised to total proteins. A representative Western blot image of OXPHOS and total proteins is shown on the right side of the figure.

Journal: Frontiers in Molecular Biosciences

Article Title: Q-Der: a next-generation CoQ10 analogue supercharging neuroprotection by combating oxidative stress and enhancing mitochondrial function

doi: 10.3389/fmolb.2025.1525103

Figure Lengend Snippet: OXPHOS protein expression. Densitometric analyses of subunits of complexes I (CI-NDUFB8, 20 kDa), II (CII-SDHB, 30 kDa), III (CIII-UQCRC2, 48 kDa) and V (CV-ATP5A, 55 kDa) of the OXPHOS system normalised to total proteins. A representative Western blot image of OXPHOS and total proteins is shown on the right side of the figure.

Article Snippet: Western blot analyses were performed using total oxidative phosphorylation (OXPHOS) cocktail from Invitrogen (458099) diluted 1:1,000 to detect individual complexes of the electron transport chain: CI-NDUFB8 (20 kDa), CII-SDHB (30 kDa), CIII-UQCRC2 (48 kDa), CIV-MTCO1 (40 kDa) and CV-ATP5A (55 kDa) ( ).

Techniques: Expressing, Western Blot