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Figure 3. NP-427 <t>downregulates</t> <t>EGFR</t> expression in the HNSCC model. (A) EGFR protein expression was determined by <t>ELISA.</t> (B) Fluorescence micro graphs of representative immunostaining of EGFR-Alexa Fluor546 conjugate in tumor tissue (red) and 4,6-diamidino-2-phenylindole dihydrochloride (DAPI) (blue) (x40), Scale bar 100 µm. (C) EGFR immunofluorescence intensity was ana lyzed by ImageJ software. Experimental groups: control group (CTR) treated with 0.9% saline solution; unloaded nanoparticles group (NP-Ø) with a concen tration of 1 mg/mL; PHT-427 inhibitor-free at concentrations of 0.5 and 1 mg/ mL group (PHT-0.5 and PHT-1, respectively) and loaded nanoparticles with the PHT-427 inhibitor at the concentrations of 0.5 and 1 mg/mL groups (NP-0.5 and NP-1, respectively) for 21 days. The diagrams for EGFR represent the mean ± SD of ng/mL for a and arbitrary units (AU) for B. p < .05 versus CTR and NP-Ø groups. (n = 32).
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Figure 3. NP-427 <t>downregulates</t> <t>EGFR</t> expression in the HNSCC model. (A) EGFR protein expression was determined by <t>ELISA.</t> (B) Fluorescence micro graphs of representative immunostaining of EGFR-Alexa Fluor546 conjugate in tumor tissue (red) and 4,6-diamidino-2-phenylindole dihydrochloride (DAPI) (blue) (x40), Scale bar 100 µm. (C) EGFR immunofluorescence intensity was ana lyzed by ImageJ software. Experimental groups: control group (CTR) treated with 0.9% saline solution; unloaded nanoparticles group (NP-Ø) with a concen tration of 1 mg/mL; PHT-427 inhibitor-free at concentrations of 0.5 and 1 mg/ mL group (PHT-0.5 and PHT-1, respectively) and loaded nanoparticles with the PHT-427 inhibitor at the concentrations of 0.5 and 1 mg/mL groups (NP-0.5 and NP-1, respectively) for 21 days. The diagrams for EGFR represent the mean ± SD of ng/mL for a and arbitrary units (AU) for B. p < .05 versus CTR and NP-Ø groups. (n = 32).
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Figure 3. NP-427 <t>downregulates</t> <t>EGFR</t> expression in the HNSCC model. (A) EGFR protein expression was determined by <t>ELISA.</t> (B) Fluorescence micro graphs of representative immunostaining of EGFR-Alexa Fluor546 conjugate in tumor tissue (red) and 4,6-diamidino-2-phenylindole dihydrochloride (DAPI) (blue) (x40), Scale bar 100 µm. (C) EGFR immunofluorescence intensity was ana lyzed by ImageJ software. Experimental groups: control group (CTR) treated with 0.9% saline solution; unloaded nanoparticles group (NP-Ø) with a concen tration of 1 mg/mL; PHT-427 inhibitor-free at concentrations of 0.5 and 1 mg/ mL group (PHT-0.5 and PHT-1, respectively) and loaded nanoparticles with the PHT-427 inhibitor at the concentrations of 0.5 and 1 mg/mL groups (NP-0.5 and NP-1, respectively) for 21 days. The diagrams for EGFR represent the mean ± SD of ng/mL for a and arbitrary units (AU) for B. p < .05 versus CTR and NP-Ø groups. (n = 32).
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Figure 3. NP-427 downregulates EGFR expression in the HNSCC model. (A) EGFR protein expression was determined by ELISA. (B) Fluorescence micro graphs of representative immunostaining of EGFR-Alexa Fluor546 conjugate in tumor tissue (red) and 4,6-diamidino-2-phenylindole dihydrochloride (DAPI) (blue) (x40), Scale bar 100 µm. (C) EGFR immunofluorescence intensity was ana lyzed by ImageJ software. Experimental groups: control group (CTR) treated with 0.9% saline solution; unloaded nanoparticles group (NP-Ø) with a concen tration of 1 mg/mL; PHT-427 inhibitor-free at concentrations of 0.5 and 1 mg/ mL group (PHT-0.5 and PHT-1, respectively) and loaded nanoparticles with the PHT-427 inhibitor at the concentrations of 0.5 and 1 mg/mL groups (NP-0.5 and NP-1, respectively) for 21 days. The diagrams for EGFR represent the mean ± SD of ng/mL for a and arbitrary units (AU) for B. p < .05 versus CTR and NP-Ø groups. (n = 32).

Journal: Drug delivery

Article Title: In vivo antitumor activity of PHT-427 inhibitor-loaded polymeric nanoparticles in head and neck squamous cell carcinoma.

doi: 10.1080/10717544.2024.2449376

Figure Lengend Snippet: Figure 3. NP-427 downregulates EGFR expression in the HNSCC model. (A) EGFR protein expression was determined by ELISA. (B) Fluorescence micro graphs of representative immunostaining of EGFR-Alexa Fluor546 conjugate in tumor tissue (red) and 4,6-diamidino-2-phenylindole dihydrochloride (DAPI) (blue) (x40), Scale bar 100 µm. (C) EGFR immunofluorescence intensity was ana lyzed by ImageJ software. Experimental groups: control group (CTR) treated with 0.9% saline solution; unloaded nanoparticles group (NP-Ø) with a concen tration of 1 mg/mL; PHT-427 inhibitor-free at concentrations of 0.5 and 1 mg/ mL group (PHT-0.5 and PHT-1, respectively) and loaded nanoparticles with the PHT-427 inhibitor at the concentrations of 0.5 and 1 mg/mL groups (NP-0.5 and NP-1, respectively) for 21 days. The diagrams for EGFR represent the mean ± SD of ng/mL for a and arbitrary units (AU) for B. p < .05 versus CTR and NP-Ø groups. (n = 32).

Article Snippet: Protein extracts were assayed by a different human enzyme-linked immunosorbent assay (elisa) kits by the following proteins: eGFR was quantified using an elisa kit (catalog number: csB-e12124h; cusabio, houston, Usa); akt(ps473) and total akt levels were measured using abcam elisa kit (catalog number: ab126433; abcam, cambridge, UK); Phospho-PDK1 (s241) and total PDK1 by the elisa Kit (catalog number: ab279889) of abcam (cambridge, UK), and human Pi3K by the elisa Kit (catalog number: csB-e08417h, cusabio, houston, Usa) according to the protocol’s instructions. the plates were assessed on the FlUOstar Omega (BMG labtech, Ortenberg, Germany) to 450 nm. these experiments were performed in triplicate for each samples (N = 31) (ctR group (n = 4); NP-Ø group (n = 5); Pht-0.5 group (n = 5); Pht-1 (n = 5); NP-0.5 (n = 6); and NP-1 group (n = 6)).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Fluorescence, Immunostaining, Immunofluorescence, Software, Control, Saline