Journal: Frontiers in Immunology

Article Title: HSP70-mediated regulation of P-glycoprotein SUMOylation suppresses ferroptosis to drive thermoresistance in non-small cell lung cancer

doi: 10.3389/fimmu.2026.1844793

Figure Lengend Snippet: HSP70 enhances P-gp protein stability by regulating SUMOylation of P-gp. (A, B) P-gp expression evaluated by RT-qPCR and western blot; (C) SUMOylation sites of P-gp predicted using the online GPS-SUMO database; (D) Interaction between P-gp and HSP70 and SUMOylation levels of P-gp assessed by Co-IP assay; (E) The SUMOylation levels of wild-type and mutant P-gp measured with Co-IP assay; (F) P-gp protein stability evaluated by CHX chase assay. All cell experiments were independently repeated three times. Data were presented as mean ± SD. Comparisons among multiple groups were performed using one-way ANOVA, followed by Tukey’s multiple comparisons test. ** P < 0.01, *** P < 0.001.

Article Snippet: Beginning in the second week after hyperthermia initiation, mice received tail vein injections of the P-gp inhibitor verapamil (25 mg/kg; HY-14275, MCE) thrice per week for three weeks , wherein verapamil was dissolved sequentially in 10% DMSO, 40% polyethylene glycol 300, 5% Tween-80, and 45% saline.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Co-Immunoprecipitation Assay, Mutagenesis

Journal: Frontiers in Immunology

Article Title: HSP70-mediated regulation of P-glycoprotein SUMOylation suppresses ferroptosis to drive thermoresistance in non-small cell lung cancer

doi: 10.3389/fimmu.2026.1844793

Figure Lengend Snippet: Knockdown of P-gp facilitates ferroptosis, which partially reverses the promoting effect of HSP70 on the resistance of NSCLC cells to hyperthermia. (A, B) RT-qPCR and western blot to evaluate P-gp expression in cells; (C) LDH to assess cell death; (D) BODIPY staining to examine lipid peroxidation; (E–G) Test kit to detect Fe 2+ , MDA, and GSH levels in cells; (H) Western blot to assess ferroptosis-related proteins (GPX4 and SLC7A11) in cells; (I) CCK-8 to examine cell viability; (J) Transwell to assess cell invasion and migration. Cell experiments were repeated thrice. Data were exhibited as mean ± SD. One-way ANOVA was utilized for data comparisons among multiple groups, and Tukey’s multiple comparison test was employed for the post hoc test. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Beginning in the second week after hyperthermia initiation, mice received tail vein injections of the P-gp inhibitor verapamil (25 mg/kg; HY-14275, MCE) thrice per week for three weeks , wherein verapamil was dissolved sequentially in 10% DMSO, 40% polyethylene glycol 300, 5% Tween-80, and 45% saline.

Techniques: Knockdown, Quantitative RT-PCR, Western Blot, Expressing, Staining, CCK-8 Assay, Migration, Comparison

Journal: Frontiers in Immunology

Article Title: HSP70-mediated regulation of P-glycoprotein SUMOylation suppresses ferroptosis to drive thermoresistance in non-small cell lung cancer

doi: 10.3389/fimmu.2026.1844793

Figure Lengend Snippet: HSP70 suppresses oxidative stress and mitochondrial dysfunction by upregulating P-gp. (A, B) Measurement of intracellular ROS and SOD levels using commercial kits; (C) Assessment of MMP assessed by JC-1 fluorescent probe; (D) Measurement of mtDNA levels; (E–H) Real-time monitoring curves of OCR in cells, along with quantitative analyses of basal respiration, ATP synthesis, and maximal respiration levels. All cell experiments were independently repeated three times. Data were presented as mean ± SD. Comparisons among groups were performed using one-way ANOVA, followed by Tukey’s multiple comparisons test. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Beginning in the second week after hyperthermia initiation, mice received tail vein injections of the P-gp inhibitor verapamil (25 mg/kg; HY-14275, MCE) thrice per week for three weeks , wherein verapamil was dissolved sequentially in 10% DMSO, 40% polyethylene glycol 300, 5% Tween-80, and 45% saline.

Techniques:

Journal: Frontiers in Immunology

Article Title: HSP70-mediated regulation of P-glycoprotein SUMOylation suppresses ferroptosis to drive thermoresistance in non-small cell lung cancer

doi: 10.3389/fimmu.2026.1844793

Figure Lengend Snippet: HSP70-mediated SUMOylation regulates P-gp to inhibit ferroptosis and thus boost resistance to hyperthermia and tumor growth in vivo . (A) Tumor images and weights; (B) Lipid peroxidation assessed by BODIPY staining; (C–F) Intratumoral ROS, Fe 2+ , MDA, and GSH levels; (G, H) Expression of ferroptosis-related proteins (GPX4 and SLC7A11), SUMO1, and SUMO2/3 detected by western blot; (I) HSP70 level in tumor tissues measured by RT-qPCR. n = 6. Data were presented as mean ± SD. Comparisons among groups were performed using one-way ANOVA, followed by Tukey’s multiple comparisons test. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Beginning in the second week after hyperthermia initiation, mice received tail vein injections of the P-gp inhibitor verapamil (25 mg/kg; HY-14275, MCE) thrice per week for three weeks , wherein verapamil was dissolved sequentially in 10% DMSO, 40% polyethylene glycol 300, 5% Tween-80, and 45% saline.

Techniques: In Vivo, Staining, Expressing, Western Blot, Quantitative RT-PCR

Journal: Frontiers in Immunology

Article Title: HSP70-mediated regulation of P-glycoprotein SUMOylation suppresses ferroptosis to drive thermoresistance in non-small cell lung cancer

doi: 10.3389/fimmu.2026.1844793

Figure Lengend Snippet: HSP70-mediated SUMOylation modulates P-gp in vivo . (A, B) IHC analysis of HSP70-positive cells and P-gp-positive cells in nude mouse tumor tissues. n = 6. Data were presented as mean ± SD. Comparisons among groups were performed using one-way ANOVA, followed by Tukey’s multiple comparisons test. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Beginning in the second week after hyperthermia initiation, mice received tail vein injections of the P-gp inhibitor verapamil (25 mg/kg; HY-14275, MCE) thrice per week for three weeks , wherein verapamil was dissolved sequentially in 10% DMSO, 40% polyethylene glycol 300, 5% Tween-80, and 45% saline.

Techniques: In Vivo

Journal: ACS Chemical Neuroscience

Article Title: Evaluation of [ 18 F]F-CNBI and [ 18 F]F-CNPI PET Probes in GSK-3β Transgenic Mice

doi: 10.1021/acschemneuro.5c00442

Figure Lengend Snippet: Time activity curves showing uptake of [ 18 F]­F-CNPI in the brain of normal, early stage and late-stage GSK-3β transgenic mice up to 60 min postinjection. (A) Comparison of uptake of [ 18 F]­F-CNPI in normal mice with and without a P-gp inhibitor. (B) Comparison of uptake of [ 18 F]­F-CNPI in early stage GSK-3β transgenic mice with and without a P-gp inhibitor. (C) Comparison of uptake of [ 18 F]­F-CNPI in late-stage GSK-3β transgenic mice with and without a P-gp inhibitor. *Adjusted P -value <0.05 with a P-gp inhibitor vs without a P-gp inhibitor, statistically significant difference. Each data point represents SUV at each frame acquired during dynamic PET imaging with a custom framing sequence of 1 × 10 s, 4 × 15 s, 8 × 30 s, 5 × 60 s, 4 × 300 s, and 3 × 600 s, representing different time points up to 60 min postinjection.

Article Snippet: The P-gp inhibitor, Tariquidar was purchased from MedChemExpress LLC, Monmouth Junction, NJ (Cat # HY-10550).

Techniques: Activity Assay, Transgenic Assay, Comparison, Imaging, Sequencing

Journal: ACS Chemical Neuroscience

Article Title: Evaluation of [ 18 F]F-CNBI and [ 18 F]F-CNPI PET Probes in GSK-3β Transgenic Mice

doi: 10.1021/acschemneuro.5c00442

Figure Lengend Snippet: Uptake of [ 18 F]­F-CNPI in the brain of normal, early stage and late-stage GSK-3β transgenic mice during the first 5 min postinjection. (A) Comparison of uptake of [ 18 F]­F-CNPI in normal mice with and without a P-gp inhibitor. (B) Comparison of uptake of [ 18 F]­F-CNPI in early stage GSK-3β transgenic mice with and without a P-gp inhibitor. (C) Comparison of uptake of [ 18 F]­F-CNPI in late-stage GSK-3β transgenic mice with and without a P-gp inhibitor. (D) Comparison of uptake of [ 18 F]­F-CNPI in normal and early stage GSK-3β transgenic mice with and without a P-gp inhibitor. (E) Comparison of uptake of [ 18 F]­F-CNPI in normal and late-stage GSK-3β transgenic mice with and without a P-gp inhibitor. (F) Comparison of uptake of [ 18 F]­F-CNPI in early stage and late-stage GSK-3β transgenic mice with and without a P-gp inhibitor. *Adjusted P -value <0.05, statistically significant difference. Each data point is an average SUV value over different frames acquired during dynamic PET imaging, representing different time points (1–5 min) postinjection.

Article Snippet: The P-gp inhibitor, Tariquidar was purchased from MedChemExpress LLC, Monmouth Junction, NJ (Cat # HY-10550).

Techniques: Transgenic Assay, Comparison, Imaging

Journal: ACS Chemical Neuroscience

Article Title: Evaluation of [ 18 F]F-CNBI and [ 18 F]F-CNPI PET Probes in GSK-3β Transgenic Mice

doi: 10.1021/acschemneuro.5c00442

Figure Lengend Snippet: Representative transverse PET/X-ray images showing uptake of [ 18 F]­F-CNBI. (A) Normal and (B) early stage GSK-3β overexpressing transgenic mice. (C–D) Time activity curves showing uptake of [ 18 F]­F-CNBI in different regions of the brain across normal and early stage GSK-3β overexpressing transgenic mice pretreated with or without a P-gp inhibitor up to 120 min postinjection. Each data point is an average SUV value over different frames acquired during dynamic PET imaging, representing different time points (5–60 min) or static PET imaging at 120 min postinjection.

Article Snippet: The P-gp inhibitor, Tariquidar was purchased from MedChemExpress LLC, Monmouth Junction, NJ (Cat # HY-10550).

Techniques: Transgenic Assay, Activity Assay, Imaging

Journal: ACS Chemical Neuroscience

Article Title: Evaluation of [ 18 F]F-CNBI and [ 18 F]F-CNPI PET Probes in GSK-3β Transgenic Mice

doi: 10.1021/acschemneuro.5c00442

Figure Lengend Snippet: Representative coronal PET/X-ray images showing uptake of [ 18 F]­F-CNPI. (A) Normal and (B) early stage GSK-3β overexpressing transgenic mice, and PET/CT images of (C) late-stage GSK-3β overexpressing transgenic mice. (D–E) Time activity graphs showing uptake of [ 18 F]­F-CNPI in different regions of the brain across normal, early stage, and late-stage GSK-3β overexpressing transgenic mice pretreated with or without a P-gp inhibitor up to 120 min postinjection. † Adjusted P -value <0.05 for early stage vs late-stage GSK-3β overexpressing transgenic groups. Adjusted P -value <0.05 for statistically significant difference. Each data point is an average SUV value over different frames acquired during dynamic PET imaging, representing different time points (5–60 min) or static PET imaging at 120 min postinjection.

Article Snippet: The P-gp inhibitor, Tariquidar was purchased from MedChemExpress LLC, Monmouth Junction, NJ (Cat # HY-10550).

Techniques: Transgenic Assay, Positron Emission Tomography-Computed Tomography, Activity Assay, Imaging

Journal: ACS Chemical Neuroscience

Article Title: Evaluation of [ 18 F]F-CNBI and [ 18 F]F-CNPI PET Probes in GSK-3β Transgenic Mice

doi: 10.1021/acschemneuro.5c00442

Figure Lengend Snippet: Box and whisker plots showing SUV of [ 18 F]­F-CNPI in different regions of the brain harvested at 120 min postinjection of [ 18 F]­F-CNPI assessed by ex vivo biodistribution analysis in (A) normal and early stage GSK-3β transgenic groups, (B) normal and late-stage GSK-3β transgenic groups, and (C) early stage and late-stage GSK-3β transgenic groups treated with or without a P-gp inhibitor. *Adjusted P -value <0.05 with a P-gp inhibitor vs without a P-gp inhibitor. Adjusted P -value <0.05 indicates a statistically significant difference.

Article Snippet: The P-gp inhibitor, Tariquidar was purchased from MedChemExpress LLC, Monmouth Junction, NJ (Cat # HY-10550).

Techniques: Whisker Assay, Ex Vivo, Transgenic Assay

Journal: ACS Chemical Neuroscience

Article Title: Evaluation of [ 18 F]F-CNBI and [ 18 F]F-CNPI PET Probes in GSK-3β Transgenic Mice

doi: 10.1021/acschemneuro.5c00442

Figure Lengend Snippet: Box and whisker plots showing SUV of [ 18 F]­F-CNBI assessed by ex vivo biodistribution analysis in (A) different regions of the brain, (B) vital organs, and (C) excretory organs harvested at 120 min postinjection of [ 18 F]­F-CNBI in normal and early stage GSK-3β transgenic groups treated with or without a P-gp inhibitor.

Article Snippet: The P-gp inhibitor, Tariquidar was purchased from MedChemExpress LLC, Monmouth Junction, NJ (Cat # HY-10550).

Techniques: Whisker Assay, Ex Vivo, Transgenic Assay