Journal: Cells

Article Title: Impact of Oncogenic Changes in p53 and KRAS on Macropinocytosis and Ferroptosis in Colon Cancer Cells and Anticancer Efficacy of Niclosamide with Differential Effects on These Two Processes

doi: 10.3390/cells13110951

Figure Lengend Snippet: Genetic background of the four human colon cancer cell lines used in the study.

Article Snippet: The p53 activator RITA (reactivation of p53 and induction of tumor cell apoptosis) was obtained from Abcam (cat. No. ab219379) (Cambridge, UK).

Techniques: Mutagenesis

Journal: Cells

Article Title: Impact of Oncogenic Changes in p53 and KRAS on Macropinocytosis and Ferroptosis in Colon Cancer Cells and Anticancer Efficacy of Niclosamide with Differential Effects on These Two Processes

doi: 10.3390/cells13110951

Figure Lengend Snippet: Expression and function of SLC38A3/SLC38A5 in the two pairs of isogenic colon cancer cell lines with and without p53 (HCT-116) and with and without G12D mutation in KRAS (SW48). Relative expression of mRNA levels as assessed by qRT-PCR ( A ). Transport function as assessed by serine uptake ( B ). Data are means ± S.E. for three independent experiments. **, p < 0.01; ***, p < 0.001. When not specified, the difference was not statistically significant.

Article Snippet: The p53 activator RITA (reactivation of p53 and induction of tumor cell apoptosis) was obtained from Abcam (cat. No. ab219379) (Cambridge, UK).

Techniques: Expressing, Mutagenesis, Quantitative RT-PCR

Journal: Cells

Article Title: Impact of Oncogenic Changes in p53 and KRAS on Macropinocytosis and Ferroptosis in Colon Cancer Cells and Anticancer Efficacy of Niclosamide with Differential Effects on These Two Processes

doi: 10.3390/cells13110951

Figure Lengend Snippet: Evidence for SLC38A5 as a transcriptional target for p53 and Myc. Locations for theoretical binding sites in human SLC38A5 gene promoter for p53 and Myc ( A ). Protein levels for p53 and Myc in the isogenic cell lines SW48 with and without KRAS-G12D mutation and HCT-116 with and without p53 ( B ). SW48 parent cells with or without treatment in the presence of the p53 activator RITA (1 μM; 16 h treatment) were used for ChIP assay to provide evidence for the binding of p53 and Myc to the SLC38A5 gene promoter ( C , D ). The ability of RITA to increase the steady-state levels of p53 was confirmed by Western blotting in all three colon cancer cell lines that expressed wild-type p53 ( E ). ***, p < 0.001.

Article Snippet: The p53 activator RITA (reactivation of p53 and induction of tumor cell apoptosis) was obtained from Abcam (cat. No. ab219379) (Cambridge, UK).

Techniques: Binding Assay, Mutagenesis, Western Blot

Journal: Cells

Article Title: Impact of Oncogenic Changes in p53 and KRAS on Macropinocytosis and Ferroptosis in Colon Cancer Cells and Anticancer Efficacy of Niclosamide with Differential Effects on These Two Processes

doi: 10.3390/cells13110951

Figure Lengend Snippet: Effect of p53 deletion and oncogenic KRAS mutation G12D on macropinocytosis. Cellular uptake of TMR-dextran was used to monitor macropinocytosis activity. The fluorescence signals were quantified as CTCF (corrected total cell fluorescence) for all four cell lines. Data are means ± S.E. ***, p < 0.001. When not specified, the difference was not statistically significant.

Article Snippet: The p53 activator RITA (reactivation of p53 and induction of tumor cell apoptosis) was obtained from Abcam (cat. No. ab219379) (Cambridge, UK).

Techniques: Mutagenesis, Activity Assay, Fluorescence

Journal: Cells

Article Title: Impact of Oncogenic Changes in p53 and KRAS on Macropinocytosis and Ferroptosis in Colon Cancer Cells and Anticancer Efficacy of Niclosamide with Differential Effects on These Two Processes

doi: 10.3390/cells13110951

Figure Lengend Snippet: Expression and function of SLC7A11 and its chaperone SLC3A2 in the two pairs of isogenic colon cancer cell lines with and without p53 (HCT-116) and with and without G12D mutation in KRAS (SW48). Relative expression of mRNA levels as assessed by qRT-PCR ( A , B ). Transport function as assessed by glutamate uptake ( C , D ). Data are means ± S.E. for three independent experiments. *, p < 0.05; ***, p < 0.001. Protein levels for SLC7A11 and SLC3A2 in the four cell lines ( E , F ).

Article Snippet: The p53 activator RITA (reactivation of p53 and induction of tumor cell apoptosis) was obtained from Abcam (cat. No. ab219379) (Cambridge, UK).

Techniques: Expressing, Mutagenesis, Quantitative RT-PCR

Journal: Cells

Article Title: Impact of Oncogenic Changes in p53 and KRAS on Macropinocytosis and Ferroptosis in Colon Cancer Cells and Anticancer Efficacy of Niclosamide with Differential Effects on These Two Processes

doi: 10.3390/cells13110951

Figure Lengend Snippet: Basal ferroptosis activity in the two pairs of isogenic colon cancer cells with and without p53 (HCT-116) and with and without G12D mutation in KRAS (SW48) as assessed by fluorescence detection of lipid radicals. Quantifications of the fluorescence signals are also given (means ± S.E. for three independent experiments). ***, p < 0.001. When not specified, the difference was not significant.

Article Snippet: The p53 activator RITA (reactivation of p53 and induction of tumor cell apoptosis) was obtained from Abcam (cat. No. ab219379) (Cambridge, UK).

Techniques: Activity Assay, Mutagenesis, Fluorescence

Journal: Cells

Article Title: Impact of Oncogenic Changes in p53 and KRAS on Macropinocytosis and Ferroptosis in Colon Cancer Cells and Anticancer Efficacy of Niclosamide with Differential Effects on These Two Processes

doi: 10.3390/cells13110951

Figure Lengend Snippet: Iron levels in the two isogenic cell lines with and without p53 (HCT-116) and with and without G12D mutation in KRAS (SW48). The fluorescence signals were also quantified (data given as means ± S.E.). **, p < 0.01; ***, p < 0.001.

Article Snippet: The p53 activator RITA (reactivation of p53 and induction of tumor cell apoptosis) was obtained from Abcam (cat. No. ab219379) (Cambridge, UK).

Techniques: Mutagenesis, Fluorescence

Journal: Cells

Article Title: Impact of Oncogenic Changes in p53 and KRAS on Macropinocytosis and Ferroptosis in Colon Cancer Cells and Anticancer Efficacy of Niclosamide with Differential Effects on These Two Processes

doi: 10.3390/cells13110951

Figure Lengend Snippet: Effects of niclosamide on intracellular pH in all four cell lines and on the transport activity of SLC38A5 in SW48 cells with G12D mutation in KRAS and on the transport activity of SLC7A11 in p53-null HCT-116 cells. Data are given as means ± S.E. *, p < 0.05; **, p < 0.01; ***, p < 0.001.

Article Snippet: The p53 activator RITA (reactivation of p53 and induction of tumor cell apoptosis) was obtained from Abcam (cat. No. ab219379) (Cambridge, UK).

Techniques: Activity Assay, Mutagenesis

Journal: Cells

Article Title: Impact of Oncogenic Changes in p53 and KRAS on Macropinocytosis and Ferroptosis in Colon Cancer Cells and Anticancer Efficacy of Niclosamide with Differential Effects on These Two Processes

doi: 10.3390/cells13110951

Figure Lengend Snippet: Induction of ferroptosis by niclosamide (5 µM) in HCT116 cells with and without p53. Quantification of the fluorescence signals (mean ± S.E.) are also given. ***, p < 0.001. When not specified, the differences were not statistically significant.

Article Snippet: The p53 activator RITA (reactivation of p53 and induction of tumor cell apoptosis) was obtained from Abcam (cat. No. ab219379) (Cambridge, UK).

Techniques: Fluorescence

Journal: Cells

Article Title: Impact of Oncogenic Changes in p53 and KRAS on Macropinocytosis and Ferroptosis in Colon Cancer Cells and Anticancer Efficacy of Niclosamide with Differential Effects on These Two Processes

doi: 10.3390/cells13110951

Figure Lengend Snippet: Blockade of niclosamide (5 µM)-induced ferroptosis by ferrostatin (10 µM) in p53 -null HCT116 cells and in SW48 cells with and without the KRAS G12D mutation.

Article Snippet: The p53 activator RITA (reactivation of p53 and induction of tumor cell apoptosis) was obtained from Abcam (cat. No. ab219379) (Cambridge, UK).

Techniques: Mutagenesis

Journal: Cells

Article Title: Impact of Oncogenic Changes in p53 and KRAS on Macropinocytosis and Ferroptosis in Colon Cancer Cells and Anticancer Efficacy of Niclosamide with Differential Effects on These Two Processes

doi: 10.3390/cells13110951

Figure Lengend Snippet: Inhibition of cell viability and proliferation by niclosamide as monitored by MTT assay in all four cell lines (HCT-116 cells with and without p53 and SW48 cells with and without KRAS-G12D mutation). Cell viability was calculated as percentage of control cells without treatment with niclosamide. Data (means ± S.E.) are from three independent experiments.

Article Snippet: The p53 activator RITA (reactivation of p53 and induction of tumor cell apoptosis) was obtained from Abcam (cat. No. ab219379) (Cambridge, UK).

Techniques: Inhibition, MTT Assay, Mutagenesis

Journal: Cells

Article Title: Impact of Oncogenic Changes in p53 and KRAS on Macropinocytosis and Ferroptosis in Colon Cancer Cells and Anticancer Efficacy of Niclosamide with Differential Effects on These Two Processes

doi: 10.3390/cells13110951

Figure Lengend Snippet: Effects of niclosamide on colony-forming ability in HCT-116 cells with and without p53 and in SW48 cells with and without KRAS-G12D mutation. Data (means ± S.E.) are from three independent experiments. *, p < 0.05; **, p < 0.01; ***, p < 0.001. When not specified, the differences were not statistically significant.

Article Snippet: The p53 activator RITA (reactivation of p53 and induction of tumor cell apoptosis) was obtained from Abcam (cat. No. ab219379) (Cambridge, UK).

Techniques: Mutagenesis

Journal: Scientific Reports

Article Title: The role of leptin in regulation of the soluble amyloid precursor protein α (sAPPα) levels in lung cancer cell media

doi: 10.1038/s41598-024-55717-y

Figure Lengend Snippet: Treatment of A549 cells with leptin increased ACh levels and inhibited the activities of p53 and AChE while opposite effects were found upon transfection with siRNA targeting either leptin or Lep-R. Cells (0.2 × 10 5 ) were grown in 10% FBS-supplemented media for 24 h. The following day, the cell monolayers were incubated in serum-free media for 24 h, then the media was replaced with fresh serum-free media. The cells were then either untreated or treated as indicated with leptin (50 ng/mL) or the indicated siRNAs and allowed to incubate for 72 h. The activity of p53 ( A ) in cell lysates, the levels of ACh in the cell media ( B ), the levels of AChE ( C ) and activity ( D ) in the media were measured as described in the “ ” section. The same amount of protein (3 µL of 600 µg/mL total protein) was used for all assays. The graphs summarize the results expressed as means ± SD (n = 5) using the GraphPad 10.0.2 software. Fold change was calculated relative to the control of each cell line ( B ) or to the A549 control ( A , C , D ). Asterisks indicate a statistically significant difference from the corresponding control, Mann–Whitney test. Statistical differences between different groups were analyzed by an ordinary one-way analysis of variance (ANOVA) followed by Tukey’s post-hoc multiple comparison test, **p < 0.01. Absence of asterisks indicates no significance.

Article Snippet: The human p53 transcription factor activity assay kit (TFEH-p53, RayBio) was used to assay the activity of p53 as we reported earlier , , .

Techniques: Transfection, Incubation, Activity Assay, Software, MANN-WHITNEY, Comparison

Journal: Journal of cell communication and signaling

Article Title: Mutant p53 gain-of-function stimulates canonical Wnt signaling via PI3K/AKT pathway in colon cancer.

doi: 10.1007/s12079-023-00793-4

Figure Lengend Snippet: Fig. 3 Mutant p53 knockdown in the SW480 cell line decreases canonical Wnt pathway. A Comparation of non-phospho- rylated β-catenin (active) and p53 protein levels measured in different colon cancer cell lines with different TP53 status, RKO (wild-type p53), SW480 (R273H p53), and SW620 (R273H p53). Densitometric analysis of immunoblots of p53 and active β-catenin was performed using GAPDH as a loading control. B Representa- tive immunoblots and graph of p53 protein in SW480 under different doses of siRNA p53 (25–100 nM); the scramble condition was used as a control (100 nM). C Luciferase activity measured with the TOP/FOP system in both scramble and siRNA p53 (100 nM) condi- tions. The relative units were normalized to Renilla activity. D Representative immunoblot and graph of c-myc normalized to GAPDH. E. Representa- tive colonies and relative area quantification corresponding to scramble and shp53 conditions of SW480-transduced cells. Graphs represent densito- metric analysis from at least three independent experiments (means ± SEM). *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: The PI3K/AKT inhibitor wortmannin was obtained from Sigma (Cat. No. 19545-26-7), and the p53 Activator III compound RITA was from Santa Cruz Biotechnology (Cat. No. sc-202743).

Techniques: Mutagenesis, Knockdown, Western Blot, Control, Luciferase, Activity Assay

Journal: Heliyon

Article Title: Heat stress suppresses MnSOD expression via p53-Sp1 interaction and induces oxidative stress damage in endothelial cells: Protective effects of MitoQ10 and Pifithrin-α

doi: 10.1016/j.heliyon.2023.e22805

Figure Lengend Snippet: Heat stress reduces cell viability, increases ROS levels, and upregulates P53 expression while downregulating MnSOD expression A) MAECs were exposed to 43 °C for 2 h, then transferred to a 37 °C incubator for rewarming. Cell viability and apoptosis were tested at different time points (1, 3, 6, and 9 h). B) Cell viability and apoptosis assay were performed after recovery for 1, 3, 6, and 9 h * p < 0.05, ** p < 0.01, *** p < 0.001. C) Mitochondrial ROS were measured using fluorescent microscope and quantitative analysis was determined using flow cytometry. Scale bar: 100 μm * p < 0.05, ** p < 0.01. D, E) The mRNA level of Tp53 and Sod2 in MAECs were detected using RT-PCR. * p < 0.05. *** p < 0.001. F) The scatter plot showed a negative correlation between the mRNA expression levels of Tp53 and Sod2 . The correlation coefficient is −0.2467 and the p value is 0.0487. G) Protein expression of p53 and MnSOD in MAECs induced by heat stress was detected using western blotting (G) and its grayscale analysis (H) was performed. I) The scatter plot displayed a negative correlation between the protein expression levels of p53 and MnSOD. The correlation coefficient is −0.5871 and the p value is 0.0006.

Article Snippet: Lentiviral particles targeting Sod2 (sc-423070-LAC, Santa Cruz), Sp1 (sc-423094-LAC, Santa Cruz), or Tp53 (sc-423509-LAC, Santa Cruz) were added to 5 μg/mL polybrene in the complete medium.

Techniques: Expressing, Apoptosis Assay, Microscopy, Flow Cytometry, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Heliyon

Article Title: Heat stress suppresses MnSOD expression via p53-Sp1 interaction and induces oxidative stress damage in endothelial cells: Protective effects of MitoQ10 and Pifithrin-α

doi: 10.1016/j.heliyon.2023.e22805

Figure Lengend Snippet: Overexpression of MnSOD alleviates ROS production and cell damage induced by heat stress A) MAECs were transfected with lentivirus targeting Sod2 , and exposed to 43 °C for 2 h, and subsequently transferred to a 37 °C incubator for rewarming. Cell viability and apoptosis assay were tested after a 6-h recovery period. B) The transfection efficiency was validated using copGFP control. Scale bar: 200 μm. C) Expression of Sod2 mRNA was measured using RT-PCR in both control and Sod2 overexpression groups under normal and heat stress conditions. ** p < 0.01, *** p < 0.001. D) Expression of MnSOD protein was measured using western blotting in both control and Sod2 overexpression groups under normal and heat stress conditions. * p < 0.05. *** p < 0.001. E, F) Cell viability and apoptosis assay were tested in both control and Sod2 overexpression groups under normal and heat stress conditions. *** p < 0.001. G) Mitochondrial ROS were visualized using fluorescent microscope and quantitative analysis was determined using flow cytometry. Scale bar: 50 μm *** p < 0.001. H) Expression of p53 protein was measured using western blotting in both control and Sod2 overexpression groups under normal and heat stress conditions with quantitative analysis performed using greyscale analysis. * p < 0.05, ** p < 0.01. I) The scatter plot indicated a negative correlation between the protein expression levels of p53 and MnSOD. The correlation coefficient is −0.7704 and the p value is < 0.0001.

Article Snippet: Lentiviral particles targeting Sod2 (sc-423070-LAC, Santa Cruz), Sp1 (sc-423094-LAC, Santa Cruz), or Tp53 (sc-423509-LAC, Santa Cruz) were added to 5 μg/mL polybrene in the complete medium.

Techniques: Over Expression, Transfection, Apoptosis Assay, Control, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Microscopy, Flow Cytometry

Journal: Heliyon

Article Title: Heat stress suppresses MnSOD expression via p53-Sp1 interaction and induces oxidative stress damage in endothelial cells: Protective effects of MitoQ10 and Pifithrin-α

doi: 10.1016/j.heliyon.2023.e22805

Figure Lengend Snippet: P53 inhibition mediates the reduction of oxidative stress injury and the increase of MnSOD level in heat stress-induced MAECs A) MAECs were transfected with siRNA targeting Tp53 , then subjected to 43 °C for 2 h, and subsequently transferred to a 37 °C incubator for rewarming. Cell viability and apoptosis assay were tested after a 6-h recovery period. B) The transfection efficiency was validated using siRNA-FITC. Scale bar: 200 μm. C) Expression of Tp5 3 mRNA was measured using RT-PCR in both control and Tp53 knockdown groups under normal and heat stress conditions. * p < 0.05, ** p < 0.01, *** p < 0.001. D) Expression of p53 protein was measured using western blotting in both control and p53 knockdown group under normal and heat stress condition (left) with quantitative analysis on the right. *** p < 0.001. E, F) Cell viability and apoptosis assay were tested in both control and Tp53 knockdown groups under normal and heat stress conditions. *** p < 0.001. G) Mitochondrial ROS level was determined using flow cytometry in both control and Tp53 knockdown groups under normal and heat stress conditions. ** p < 0.01, *** p < 0.001. H) Expression of Sod2 mRNA was measured using RT-PCR in both control and Tp53 knockdown groups under normal and heat stress conditions. * p < 0.05, ** p < 0.01, *** p < 0.001. I) Expression of p53 protein was measured using western blotting in both control and Tp53 knockdown groups under normal and heat stress conditions with quantitative results performed using greyscale analysis. ** p < 0.01, *** p < 0.001.

Article Snippet: Lentiviral particles targeting Sod2 (sc-423070-LAC, Santa Cruz), Sp1 (sc-423094-LAC, Santa Cruz), or Tp53 (sc-423509-LAC, Santa Cruz) were added to 5 μg/mL polybrene in the complete medium.

Techniques: Inhibition, Transfection, Apoptosis Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Control, Knockdown, Western Blot, Flow Cytometry

Journal: Heliyon

Article Title: Heat stress suppresses MnSOD expression via p53-Sp1 interaction and induces oxidative stress damage in endothelial cells: Protective effects of MitoQ10 and Pifithrin-α

doi: 10.1016/j.heliyon.2023.e22805

Figure Lengend Snippet: Inhibition of p53 using Pifithrin attenuated mitochondrial ROS and alleviated MnSOD reduction induced by heat stress A) Mice were pre-treated with Pifithrin-α (3 mg/kg/day) i. p. for 3 d prior to heat stress treatment. B) Rectal temperature was measured in both control and Pifithrin-α treated groups under normal and heat stress conditions. *** p < 0.001. n = 6 in each group. C) Expression of p53 protein was measured using western blotting in both control and Pifithrin-α treated groups under normal and heat stress conditions with quantitative results performed using greyscale analysis. ** p < 0.01, *** p < 0.001. D) Mitochondrial ROS level was determined using flow cytometry in both control and Pifithrin-α treated groups under normal and heat stress conditions. ** p < 0.01, *** p < 0.001. E) Plasma ROS was evaluated using a microplate reader in both control and Pifithrin-α treated groups under normal and heat stress conditions. * p < 0.05, ** p < 0.01. F) Expression of MnSOD protein was measured using western blotting in both control and Pifithrin-α treated groups under normal and heat stress conditions. ** p < 0.01, *** p < 0.001. G) The scatter plot indicated a negative correlation between the protein expression levels of p53 and MnSOD. The correlation coefficient is −0.6145 and the p value is 0.0015.

Article Snippet: Lentiviral particles targeting Sod2 (sc-423070-LAC, Santa Cruz), Sp1 (sc-423094-LAC, Santa Cruz), or Tp53 (sc-423509-LAC, Santa Cruz) were added to 5 μg/mL polybrene in the complete medium.

Techniques: Inhibition, Control, Expressing, Western Blot, Flow Cytometry, Clinical Proteomics

Journal: Heliyon

Article Title: Heat stress suppresses MnSOD expression via p53-Sp1 interaction and induces oxidative stress damage in endothelial cells: Protective effects of MitoQ10 and Pifithrin-α

doi: 10.1016/j.heliyon.2023.e22805

Figure Lengend Snippet: Oxidative stress Injury and MnSOD Reduction induced by p53 overexpression in MAECs is not exacerbated under heat stress A) MAECs were transfected with lentivirus particles targeting Tp53 prior to heat stress treatment. B) Transfection efficiency of lentiviral particles targeting Tp53 was validated with copGFP control. Scale bar: 200 μm. C) The protein overexpression of p53 after transfection was measured using western blotting. ** p < 0.01. D, E) Cell viability and apoptosis assay were tested in both control and Tp53 overexpression groups under normal and heat stress conditions. *** p < 0.001. F) Mitochondrial ROS level was determined using flow cytometry in both control and Tp53 overexpression groups under normal and heat stress conditions. * p < 0.05, *** p < 0.001. G) Expression of MnSOD protein was measured using western blotting in both control and Tp53 overexpression groups under normal and heat stress conditions. ** p < 0.01, *** p < 0.001.

Article Snippet: Lentiviral particles targeting Sod2 (sc-423070-LAC, Santa Cruz), Sp1 (sc-423094-LAC, Santa Cruz), or Tp53 (sc-423509-LAC, Santa Cruz) were added to 5 μg/mL polybrene in the complete medium.

Techniques: Over Expression, Transfection, Control, Western Blot, Apoptosis Assay, Flow Cytometry, Expressing

Journal: Heliyon

Article Title: Heat stress suppresses MnSOD expression via p53-Sp1 interaction and induces oxidative stress damage in endothelial cells: Protective effects of MitoQ10 and Pifithrin-α

doi: 10.1016/j.heliyon.2023.e22805

Figure Lengend Snippet: Heat stress activates p53 and inhibits SP1 to downregulate MnSOD expression in mouse endothelial cells A) MAECs were transfected with siRNA targeting Sp1 and Tp53 , or lentivirus particles targeting Sp1 prior to heat stress treatment. B) The transfection efficiency was validated using siRNA-FITC. Scale bar: 200 μm. C) The expression of Sp1 protein in both Sp1 knockdown and overexpression groups using western blotting. * p < 0.05, ** p < 0.01, *** p < 0.001. D) The scatter plot showed a positive correlation between the protein expression levels of Sp1 and MnSOD. The correlation coefficient is 0.8511 and p -value is < 0.0001. E, F) Cell viability and apoptosis assay were tested in control, siRNA- Tp53 , SP1 overexpression, and siRNA- Tp53-Sp1 groups under heat stress conditions. ** p < 0.01, *** p < 0.001. G) Mitochondrial ROS level was determined using flow cytometry in control, siRNA- Tp53 , SP1 overexpression, and siRNA- Tp53-Sp1 groups under heat stress conditions. * p < 0.05, ** p < 0.01. H) Mice were treated with MitoQ10 (500 μM/day) or saline 3 weeks prior to heat stress. I) Rectal temperature was measured in both control and MitoQ10 treated groups under normal and heat stress conditions. *** p < 0.001. n = 6 in each group. J) Mitochondrial ROS level was determined using flow cytometry in both control and MitoQ10 treated groups under normal and heat stress conditions. ** p < 0.01, *** p < 0.001. K) The measurement of Plasma ROS was conducted by a microplate reader in both control and MitoQ10 treated groups under normal and heat stress conditions. * p < 0.05, ** p < 0.01. L): P53 interacts with Sp1 in co-IP assays in MAECs incubated under heat stress conditions.

Article Snippet: Lentiviral particles targeting Sod2 (sc-423070-LAC, Santa Cruz), Sp1 (sc-423094-LAC, Santa Cruz), or Tp53 (sc-423509-LAC, Santa Cruz) were added to 5 μg/mL polybrene in the complete medium.

Techniques: Expressing, Transfection, Knockdown, Over Expression, Western Blot, Apoptosis Assay, Control, Flow Cytometry, Saline, Clinical Proteomics, Co-Immunoprecipitation Assay, Incubation