Journal: iScience

Article Title: Targeted PGC-1α gene delivery by GV-assisted ultrasound cavitation for renal ischemia-reperfusion injury therapy

doi: 10.1016/j.isci.2025.114374

Figure Lengend Snippet: Expression of PGC-1α and P-selectin in the kidneys after renal I/R (A and B) Protein expression levels of PGC-1α in the kidneys of mice with acute renal I/R; in RMVECs after H/R, were evaluated by Western blot. (C) The expression levels of PGC-1α in the ECs of kidneys in mice with acute renal I/R were observed by immunofluorescence (scale bars: 20 μm). (D) Protein expression levels of ICAM-1, E-selectin, and P-selectin in the kidneys of mice with acute renal I/R were evaluated by Western blot. (E) The expression levels of P-selectin in the ECs of kidneys in mice with acute renal I/R were observed by immunofluorescence (scale bars: 20 μm). (F) Quantification of CD31 + /P-selectin + cells in renal immunofluorescence images. (G) Protein expression levels of ICAM-1 and P-selectin in RMVECs after H/R were evaluated by Western blot. Protein bands were quantitatively analyzed by a histogram. Data are expressed as the mean ± standard deviation (SD) (A, B, D, and G: n = 3, n represents the number of independent experiments; F: n = 3, n represents the number of mice in each group). t test was utilized for statistical analysis (∗∗ p < 0.01, ∗∗∗ p < 0.001).

Article Snippet: P-selectin Antibody , Proteintech , Cat # 60322-1-Ig; RRID: AB_2881433.

Techniques: Expressing, Western Blot, Immunofluorescence, Standard Deviation

Journal: iScience

Article Title: Targeted PGC-1α gene delivery by GV-assisted ultrasound cavitation for renal ischemia-reperfusion injury therapy

doi: 10.1016/j.isci.2025.114374

Figure Lengend Snippet: Renal EC targeting capacity of Fuc-pPGC-1α-GVs (A) FITC-labeled Fuc-pPGC-1α-GVs were incubated with H/R injured RMVECs in vitro , and the internalization of Fuc-pPGC-1α-GVs was imaged with IVIS spectrum. (B) The amount of Fuc-pPGC-1α-GVs internalized was quantitatively analyzed by histogram. (C and D) RMVECs with the high expression of P-selectin induced by H/R showed enhanced internalization of Fuc-pPGC-1α-GVs in vitro (scale bars: 50 μm). Fluorescence intensity was quantitatively analyzed by a histogram. (E) Organ distributions of Dex-pPGC-1α-GVs and Fuc-pPGC-1α-GVs in mice were detected by FITC radiant efficiency after a single intravenous injection of Dex-pPGC-1α-GVs or Fuc-pPGC-1α-GVs. The histogram is for the quantitative analysis of Fuc-pPGC-1α-GVs accumulated in the injured kidney by FITC signals (left) or by kidney-to-liver fluorescence intensity ratio (right) at 1, 3, 6, 12, and 24 h after the injection of Fuc-pPGC-1α-GVs. The radiant efficiency of FITC was expressed as [photons/s/cm 2 /steradian]/[μW cm −2 ]. Lu: lung, H: heart, Li: liver, S: spleen, K: kidney. #1: Normal mice were given a single intravenous injection of Fuc-pPGC-1α-GVs; #2: Mice with renal IRI were given a single intravenous injection of Dex-pPGC-1α-GVs; #3: Mice with renal IRI were given a single intravenous injection of Fuc-pPGC-1α-GVs. Data are expressed as the mean ± SD (B and C: n = 3, n represents the number of independent experiments; E: n = 3, n represents the number of mice in each group). t test was utilized for statistical analysis (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001).

Article Snippet: P-selectin Antibody , Proteintech , Cat # 60322-1-Ig; RRID: AB_2881433.

Techniques: Labeling, Incubation, In Vitro, Expressing, Fluorescence, Injection

Journal: iScience

Article Title: Targeted PGC-1α gene delivery by GV-assisted ultrasound cavitation for renal ischemia-reperfusion injury therapy

doi: 10.1016/j.isci.2025.114374

Figure Lengend Snippet: Expression of PGC-1α and P-selectin in the kidneys after renal I/R (A and B) Protein expression levels of PGC-1α in the kidneys of mice with acute renal I/R; in RMVECs after H/R, were evaluated by Western blot. (C) The expression levels of PGC-1α in the ECs of kidneys in mice with acute renal I/R were observed by immunofluorescence (scale bars: 20 μm). (D) Protein expression levels of ICAM-1, E-selectin, and P-selectin in the kidneys of mice with acute renal I/R were evaluated by Western blot. (E) The expression levels of P-selectin in the ECs of kidneys in mice with acute renal I/R were observed by immunofluorescence (scale bars: 20 μm). (F) Quantification of CD31 + /P-selectin + cells in renal immunofluorescence images. (G) Protein expression levels of ICAM-1 and P-selectin in RMVECs after H/R were evaluated by Western blot. Protein bands were quantitatively analyzed by a histogram. Data are expressed as the mean ± standard deviation (SD) (A, B, D, and G: n = 3, n represents the number of independent experiments; F: n = 3, n represents the number of mice in each group). t test was utilized for statistical analysis (∗∗ p < 0.01, ∗∗∗ p < 0.001).

Article Snippet: After blocking with 10% goat serum, cell samples were incubated with primary antibodies against P-selectin (1:200; 60322, Proteintech), followed by secondary antibodies labeled Alexa Fluor 647 (1:200; BD9011, AbBox).

Techniques: Expressing, Western Blot, Immunofluorescence, Standard Deviation

Journal: iScience

Article Title: Targeted PGC-1α gene delivery by GV-assisted ultrasound cavitation for renal ischemia-reperfusion injury therapy

doi: 10.1016/j.isci.2025.114374

Figure Lengend Snippet: Renal EC targeting capacity of Fuc-pPGC-1α-GVs (A) FITC-labeled Fuc-pPGC-1α-GVs were incubated with H/R injured RMVECs in vitro , and the internalization of Fuc-pPGC-1α-GVs was imaged with IVIS spectrum. (B) The amount of Fuc-pPGC-1α-GVs internalized was quantitatively analyzed by histogram. (C and D) RMVECs with the high expression of P-selectin induced by H/R showed enhanced internalization of Fuc-pPGC-1α-GVs in vitro (scale bars: 50 μm). Fluorescence intensity was quantitatively analyzed by a histogram. (E) Organ distributions of Dex-pPGC-1α-GVs and Fuc-pPGC-1α-GVs in mice were detected by FITC radiant efficiency after a single intravenous injection of Dex-pPGC-1α-GVs or Fuc-pPGC-1α-GVs. The histogram is for the quantitative analysis of Fuc-pPGC-1α-GVs accumulated in the injured kidney by FITC signals (left) or by kidney-to-liver fluorescence intensity ratio (right) at 1, 3, 6, 12, and 24 h after the injection of Fuc-pPGC-1α-GVs. The radiant efficiency of FITC was expressed as [photons/s/cm 2 /steradian]/[μW cm −2 ]. Lu: lung, H: heart, Li: liver, S: spleen, K: kidney. #1: Normal mice were given a single intravenous injection of Fuc-pPGC-1α-GVs; #2: Mice with renal IRI were given a single intravenous injection of Dex-pPGC-1α-GVs; #3: Mice with renal IRI were given a single intravenous injection of Fuc-pPGC-1α-GVs. Data are expressed as the mean ± SD (B and C: n = 3, n represents the number of independent experiments; E: n = 3, n represents the number of mice in each group). t test was utilized for statistical analysis (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001).

Article Snippet: After blocking with 10% goat serum, cell samples were incubated with primary antibodies against P-selectin (1:200; 60322, Proteintech), followed by secondary antibodies labeled Alexa Fluor 647 (1:200; BD9011, AbBox).

Techniques: Labeling, Incubation, In Vitro, Expressing, Fluorescence, Injection