Journal: Small (Weinheim an der Bergstrasse, Germany)

Article Title: Capsaicin Enhanced the Efficacy of Photodynamic Therapy Against Osteosarcoma via a Pro-Death Strategy by Inducing Ferroptosis and Alleviating Hypoxia.

doi: 10.1002/smll.202306916

Figure Lengend Snippet: Figure 3. Responsive activation of TRPV1-mediated intracellular Ca2+ concentration. A) CLSM images of 143B and HOS cells in various groups after using the Fluo-4 probe to monitor Ca2+ influx. The scale bar is 20 μm. B,C) FC quantitative value of Ca2+ uptake in 143B and HOS cells after different treatments. D) The protein expression level of TRPV1 was determined by western blot analysis. E) Statistical analyses of TRPV1 (the data are presented as the mean ± SD, n = 3, ***p < 0.001).

Article Snippet: Mouse anti-human TRPV1 (cat. 66983-1-Ig), mouse anti-human NQO1 (cat. 67240-1-Ig), rabbit anti-human GCLM (cat. 14241-1-AP), rabbit anti-human PCNA (cat. 10205-2-AP), rabbit anti-human KI67 (cat. 27309-1-AP), and rabbit antihuman HIF-1α (cat. 20960-1-AP) were acquired from Proteintech Corp (USA).

Techniques: Activation Assay, Concentration Assay, Expressing, Western Blot

Journal: Small (Weinheim an der Bergstrasse, Germany)

Article Title: Capsaicin Enhanced the Efficacy of Photodynamic Therapy Against Osteosarcoma via a Pro-Death Strategy by Inducing Ferroptosis and Alleviating Hypoxia.

doi: 10.1002/smll.202306916

Figure Lengend Snippet: Figure 4. Evaluation of ferroptosis. The production of Lipid-ROS in A) 143B and B) HOS cells after different interventions as measured by CLSM. The scale bars represent 20 μm. C) FC analysis of Lipid-ROS after different interventions in 143B and HOS cells. D,E) Confocal images and FC analyses of ROS generation in 143B and HOS cells treated with different treatments and stained with DCFH-DA (scale bar = 20 μm). F,G) Quantitative value of intracellular C11-BODIPY and DCFH-DA FL intensity (the data are presented as the mean ± SD, n = 3, ***p < 0.001). H) The morphology of mitochondria after various treatments as observed by TEM. The scale bars represent 2 μm. I) The expression level of Gpx4 protein was measured by western blot analysis. J) The expression of TRPV1 and Gpx4 by IF observation after different treatments in 143B cells (scale bar = 100 μm).

Article Snippet: Mouse anti-human TRPV1 (cat. 66983-1-Ig), mouse anti-human NQO1 (cat. 67240-1-Ig), rabbit anti-human GCLM (cat. 14241-1-AP), rabbit anti-human PCNA (cat. 10205-2-AP), rabbit anti-human KI67 (cat. 27309-1-AP), and rabbit antihuman HIF-1α (cat. 20960-1-AP) were acquired from Proteintech Corp (USA).

Techniques: Staining, Expressing, Western Blot

Journal: Small (Weinheim an der Bergstrasse, Germany)

Article Title: Capsaicin Enhanced the Efficacy of Photodynamic Therapy Against Osteosarcoma via a Pro-Death Strategy by Inducing Ferroptosis and Alleviating Hypoxia.

doi: 10.1002/smll.202306916

Figure Lengend Snippet: Figure 5. A) In MG63, HOS, and 143B cells, the effects of CAP on the intracellular oxygen level after different treatments were observed and analyzed by confocal microscopy (scale bar = 20 μm) and FC. B) Quantitative value of intracellular Image-iT Green Hypoxia Reagent FL intensity (the data are presented as the mean ± SD, n = 3, ***p < 0.001). C,D) The expression level of HIF-1𝛼protein was analyzed by western blot (the data are presented as the mean ± SD, n = 3, ***p < 0.001). E,F) The expression of TRPV1 and HIF-1𝛼by IF observation after different treatments in 143B and HOS cells (scale bar = 100 μm).

Article Snippet: Mouse anti-human TRPV1 (cat. 66983-1-Ig), mouse anti-human NQO1 (cat. 67240-1-Ig), rabbit anti-human GCLM (cat. 14241-1-AP), rabbit anti-human PCNA (cat. 10205-2-AP), rabbit anti-human KI67 (cat. 27309-1-AP), and rabbit antihuman HIF-1α (cat. 20960-1-AP) were acquired from Proteintech Corp (USA).

Techniques: Confocal Microscopy, Expressing, Western Blot

Journal: Small (Weinheim an der Bergstrasse, Germany)

Article Title: Capsaicin Enhanced the Efficacy of Photodynamic Therapy Against Osteosarcoma via a Pro-Death Strategy by Inducing Ferroptosis and Alleviating Hypoxia.

doi: 10.1002/smll.202306916

Figure Lengend Snippet: Figure 8. Evaluation and biosafety of different treatments. A) H&E staining and images of PCNA, Ki67, TRPV1, Gpx4, and HIF-1𝛼. IHC staining in tumor specimens after various treatments (scale bar = 100 μm). B–F) IOD values in various groups were analyzed by processing IHC staining through ImagePro Plus. G) Triple IF staining of TRPV1 (red), Gpx4 (orange), and HIF-1𝛼(green) in tumor specimens after various treatments (scale bar = 50 μm). H) TUNEL staining of tumor sections after various treatments (scale bar = 50 μm). I) H&E staining of major organs of mice after different treatments (scale bar = 100 μm). J) Hematological and blood biochemical tests of mice after various treatments.

Article Snippet: Mouse anti-human TRPV1 (cat. 66983-1-Ig), mouse anti-human NQO1 (cat. 67240-1-Ig), rabbit anti-human GCLM (cat. 14241-1-AP), rabbit anti-human PCNA (cat. 10205-2-AP), rabbit anti-human KI67 (cat. 27309-1-AP), and rabbit antihuman HIF-1α (cat. 20960-1-AP) were acquired from Proteintech Corp (USA).

Techniques: Staining, Immunohistochemistry, TUNEL Assay

Journal: Journal of Inflammation Research

Article Title: Characterization of Early Inflammatory Events Leading to Provoked Vulvodynia Development in Rats

doi: 10.2147/jir.s367193

Figure Lengend Snippet: Figure 11 Long-term effects of mast cells stabilizer KF on TRPV1 and TRPA1 expression in vulvar nerves after 81 days of the 3rd round of zymosan/saline administration. (A) The expression of TRPV1 channel in vulva nerves, 81 days after the 3rd zymosan/saline administration. Coexpression of TRPV1 channel (green) and neuronal PGP 9.5 (red; white arrows) merged with dapi stain (blue). (B) The expression of TRPA1 channel in vulva nerves, 81 days after the 3rd round zymosan\saline administration. Coexpression of TRPA1 channel (red) and neuronal PGP 9.5 (green; white arrows) merged with dapi stain (blue). Scale bar: 50 µm. (C) Fluorescence intensity (arbitrary units) of TRPV1, TRPA1, TRPV1/PGP-9.5, and TRPA1/PGP-9.5 in the Saline- ketotifen, the zymosan- ketotifen, and the zymosan-saline group 81 of the 3rd round (n=7 per group). Mean ± SEM. *P<0.05, ***P<0.001.

Article Snippet: Afterward, to remove background staining, we used Background Buster (#NB306-50, INNOVEX), followed by three cycles of wash buffer for twominutes each, then incubated for one hour at room temperature in a humidity chamber with one of the first antibodies: anti-rat TRPV1 channel ATTO-488 (1:240, cat.ACC-029-AG, Alomone labs, Jerusalem), anti-rat TRPA1 channel (1:240, cat.ACC-037, Alomone labs, Jerusalem), anti-rat protein gene product 9.5 (PGP 9.5, 1:500, NB300-676, Novus Biological), rabbit antiChymase (1:250, NBP2-257441, Novus Biological) and mouse anti-Tryptase (1:250, NBP2-26444, Novus Biological).

Techniques: Expressing, Saline, Staining, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Platinum-Based Drugs Cause Mitochondrial Dysfunction in Cultured Dorsal Root Ganglion Neurons

doi: 10.3390/ijms21228636

Figure Lengend Snippet: Cytosolic and mitochondrial calcium concentration of TRPA1- or TRPV1-positive DRG neurons during exposure to 10 µM cisplatin. ( A ) Immunostaining of transient receptor potential ankyrin 1 (TRPA1)- or transient receptor potential vanilloid 1 (TRPV1)-positive DRG neurons and live imaging of Fluo-4 (green) and Rhod-2 (red). ( B ) The cytosolic calcium of TRPV1-positive DRG neurons concentration instantly increased to 1.067 ± 0.018 (* p < 0.05) after 5 min and continuously increased to 1.394 ± 0.068 (** p < 0.01) after 70 min. The mitochondrial calcium concentration declined to 1.095 ± 0.009 (* p < 0.05) after 5 min for TRPV1-positive DRG neurons and steadily declined to 0.577 ± 0.044 after 70 min (** p < 0.01). ( C ) During the exposure of 10 µM cisplatin, the cytosolic calcium concentration of TRPA1-positive neurons increased after 5 min to 1.024 ± 0.011 (** p < 0.01) until 70 min at 1.240 ± 0.074 (*** p < 0.001). The mitochondrial calcium concentration was lower after 10 min at 0.928 ± 0.029 (*** p < 0.001) until it reached 0.790 ± 0.050 at 70 min (*** p < 0.001). ( D ) The cytosolic calcium concentration of TRPV1-positive DRG neurons was increased after 15 min to 65 min compared to TRPA1-positive DRG neurons (* p < 0.05, ** p < 0.01). ( E ) The mitochondrial calcium concentration of TRPA1- or TRPV1-positive DRG neurons was different after 5 min until the end of the experiment (* p < 0.05, ** p < 0.01). TRPV1-positive sensory neurons had a lower mitochondrial calcium concentration during exposure to 10 µM cisplatin after 15 min (* p < 0.05, ** p < 0.01). n = 6 cells per condition. * = significant effect between TRPA1+ and TRPV1+ DRG neurons. Scale = 50 µm.

Article Snippet: Anti-TRPV1 (extracellular) , 1:200, rabbit , Alomone Labs, Israel.

Techniques: Concentration Assay, Immunostaining, Imaging

Journal: International Journal of Molecular Sciences

Article Title: Platinum-Based Drugs Cause Mitochondrial Dysfunction in Cultured Dorsal Root Ganglion Neurons

doi: 10.3390/ijms21228636

Figure Lengend Snippet: Cytosolic and mitochondrial calcium concentration of TRPA1- or TRPV1-positive DRG neurons during exposure to 10 µM oxaliplatin. ( A ) Immunostaining of TRPA1- or TRPV1-positive DRG neurons and live imaging of Fluo-4 (green) and Rhod-2 (red) before and after application of 10 µM cisplatin or oxaliplatin. ( B ) TRPA1-positive DRG neurons showed an immediate increase of the cytosolic calcium concentration to 1.037 ± 0.006 (** p < 0.01) after 5 min and continuously increased to 1.372 ± 0.062 after 70 min (*** p < 0.001). The mitochondrial calcium concentration decreased after 5 min to 0.967 ± 0.008 (*** p < 0.001) and declined to 0.712 ± 0.010 after 70 min (*** p < 0.001). ( C ) The TRPV1-positive DRG neurons showed an immediate increase of the cytosolic calcium concentration to 1.049 ± 0.005 (*** p < 0.001) after 5 min and increased to 1.480 ± 0.034 after 70 min (*** p < 0.001). The mitochondrial calcium concentration decreased to 0.959 ± 0.020 (* p < 0.05) after 5 min and to 0.725 ± 0.077 (** p < 0.01) after 70 min (* p < 0.05). ( D ) No difference could be determined in the cytosolic calcium concentration of TRPA1- or TRPV1-positive DRG neurons ( p > 0.05). ( E ) No difference could be determined in the mitochondrial calcium concentration of TRPA1- or TRPV1-positive DRG neurons ( p > 0.05). n = 6 cells per condition. * = significant effect between TRPA1+ and TRPV1+ DRG neurons. Scale = 50 µm.

Article Snippet: Anti-TRPV1 (extracellular) , 1:200, rabbit , Alomone Labs, Israel.

Techniques: Concentration Assay, Immunostaining, Imaging

Journal: International Journal of Molecular Sciences

Article Title: Platinum-Based Drugs Cause Mitochondrial Dysfunction in Cultured Dorsal Root Ganglion Neurons

doi: 10.3390/ijms21228636

Figure Lengend Snippet: Relative cytosolic and mitochondrial calcium concentration during exposure to 10 µM cis- or oxaliplatin of TRPA1- or TRPV1-positive DRG neurons. ( A ) No differences in the cytosolic calcium concentration of TRPA1-positive DRG neurons after exposure to 10 µM cis- or oxaliplatin could be determined ( p > 0.05). ( B ) The cytosolic calcium of TRPV1-positive DRG neurons concentration during exposure to cis- or oxaliplatin showed no difference ( p > 0.05). ( C ) During exposure to 10 µM cis- or oxaliplatin, no difference in the mitochondrial calcium concentration of TRPA1-positive DRG neurons could be determined ( p > 0.05). ( D ) While exposing TRPV1-positive DRG neurons, differences between the two chemotherapeutics on the mitochondrial calcium concentration could be determined after 20 min. The mitochondrial calcium concentration of TRPV1-positive DRG neurons exposed to oxaliplatin was higher at 0.856 ± 0.033 compared to 0.727 ± 0.040 (* p < 0.05). The mitochondrial calcium concentration was higher for oxaliplatin-exposed TRPV1-positive DRG neurons until 0.786 ± 0.058 after 40 min (* p < 0.05). n = 6 cells per condition. * = significant effect between cis- and oxaliplatin, ** p < 0.01

Article Snippet: Anti-TRPV1 (extracellular) , 1:200, rabbit , Alomone Labs, Israel.

Techniques: Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Platinum-Based Drugs Cause Mitochondrial Dysfunction in Cultured Dorsal Root Ganglion Neurons

doi: 10.3390/ijms21228636

Figure Lengend Snippet: ROS production of TRPA1- or TRPV1-positive DRG neurons during exposure to 10 µM cisplatin. ( A ) Fluorescence of CellRox (green) in TRPA1- or TRPV1-positive DRG neurons during exposure to 10 µM cisplatin at different time points. ( B ) TRPA1-positive sensory neurons showed an instant increase of the ROS production to 1.431 ± 0.042 (*** p < 0.001) after 5 min and reached its peak at 2.100 ± 0.197 after 30 min (*** p < 0.001). After 50 min, the ROS production dropped below the control level at 50 min to 0.607 ± 0.036 (*** p < 0.001) and further declined to 0.370 ± 0.035 (*** p < 0.001) after 70 min. ( C ) After exposure to 10 µM cisplatin of the TRPV1-positive DRG neurons, the ROS production increased to 1.238 ± 0.042 (*** p < 0.001) after 5 min and dropped below control level at 50 min at 0.873 ± 0.068 (* p < 0.05) and steadily declined to 0.756 ± 0.072 (** p < 0.01) after 70 min. n = 6 cells per condition. * = significant effect compared to control level. Scale = 50 µm.

Article Snippet: Anti-TRPV1 (extracellular) , 1:200, rabbit , Alomone Labs, Israel.

Techniques: Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Platinum-Based Drugs Cause Mitochondrial Dysfunction in Cultured Dorsal Root Ganglion Neurons

doi: 10.3390/ijms21228636

Figure Lengend Snippet: ROS production of TRPA1- or TRPV1-positive DRG neurons during exposure to 10 µM oxaliplatin. ( A ) Fluorescence of CellRox (green) of TRPA1- or TRPV1-positive DRG neurons at different time points. ( B ) The ROS production of TRPA1-positive DRG neurons increased after 5 min to 1.421 ± 0.103 (** p < 0.01) during exposure to 10 µM oxaliplatin and dropped to control level after 40 min at 1.31 ± 0.194 ( p > 0.05). ( C ) The TRPV1-positive sensory neurons had an increased ROS production after 5 min at 1.080 ± 0.020 (** p < 0.01) and dropped to the control level after 10 min at 1.033 ± 0.036 ( p > 0.05). After 30 min, the ROS production declined below the control level at 0.810 ± 0.079 (* p < 0.05) and steadily declined to 0.429 ± 0.050 after 70 min (*** p < 0.001). n = 6 cells per condition. * = significant effect compared to control level. Scale = 20 µm.

Article Snippet: Anti-TRPV1 (extracellular) , 1:200, rabbit , Alomone Labs, Israel.

Techniques: Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Platinum-Based Drugs Cause Mitochondrial Dysfunction in Cultured Dorsal Root Ganglion Neurons

doi: 10.3390/ijms21228636

Figure Lengend Snippet: The ROS production of TRPA1- or TRPV1-positive DRG neurons during exposure to cis- or oxaliplatin. ( A ) A difference could be determined for TRPA1-positive DRG neurons at 30 min with a higher ROS production for cisplatin-exposed DRG neurons at 2.100 ± 0.197 (* p < 0.05). After 45 min (1.194 ± 0.223; * p < 0.05) until 70 min (1.022 ± 0.233; * p < 0.05), the ROS production was higher for oxaliplatin-exposed neurons. ( B ) The ROS production of TRPV1-positive DRG neurons was higher for cisplatin-exposed neurons over the entire experimental period (* p < 0.05, ** p < 0.01). n = 6 cells per condition. * = significant effect compared to control level.

Article Snippet: Anti-TRPV1 (extracellular) , 1:200, rabbit , Alomone Labs, Israel.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Platinum-Based Drugs Cause Mitochondrial Dysfunction in Cultured Dorsal Root Ganglion Neurons

doi: 10.3390/ijms21228636

Figure Lengend Snippet: Antibodies and dilution used for the TRPA1- and TRPV1-positive staining.

Article Snippet: Anti-TRPV1 (extracellular) , 1:200, rabbit , Alomone Labs, Israel.

Techniques: Staining

Journal: Investigative and Clinical Urology

Article Title: Changes in transient receptor potential vanilloid 1 and transient receptor potential vanilloid 4 in patients with lower urinary tract dysfunction

doi: 10.4111/icu.20210418

Figure Lengend Snippet: Comparisons of TRPV1 and TRPV4 expression levels (A) between the DU group and non-DU group and (B) between the urgency group and non-urgency group. TRPV, transient receptor potential vanilloid; DU, detrusor underactivity. * Significant difference between the two groups (p<0.05).

Article Snippet: After two freeze-thaw cycles were performed, the homogenate was centrifuged at 2°C to 8°C for 5 minutes at 5,000× g . The levels of TRPV1 and TRPV4 in the specimen were measured according to the manufacturer’s instructions using a human TRPV1 ELISA kit and a human TRPV4 ELISA kit (Cusabio Biotech Co., Houston, TX, USA).

Techniques: Expressing

Journal: Investigative and Clinical Urology

Article Title: Changes in transient receptor potential vanilloid 1 and transient receptor potential vanilloid 4 in patients with lower urinary tract dysfunction

doi: 10.4111/icu.20210418

Figure Lengend Snippet: Correlation of TRPV1 and TRPV4 with urodynamic parameters

Article Snippet: After two freeze-thaw cycles were performed, the homogenate was centrifuged at 2°C to 8°C for 5 minutes at 5,000× g . The levels of TRPV1 and TRPV4 in the specimen were measured according to the manufacturer’s instructions using a human TRPV1 ELISA kit and a human TRPV4 ELISA kit (Cusabio Biotech Co., Houston, TX, USA).

Techniques:

Journal: Investigative and Clinical Urology

Article Title: Changes in transient receptor potential vanilloid 1 and transient receptor potential vanilloid 4 in patients with lower urinary tract dysfunction

doi: 10.4111/icu.20210418

Figure Lengend Snippet: Immunofluorescence staining of TRPV1 and TRPV4. (A) TRPV1 in the DU group. (B) TRPV1 in the non-DU group. (C) TRPV4 in the DU group. (D) TRPV4 in the non-DU group. (E) TRPV1 in the urgency group. (F) TRPV1 in the non-urgency group. (G) TRPV4 in the urgency group. (H) TRPV4 in the non-urgency group. Scale bars: 200 µm. TRPV, transient receptor potential vanilloid; DU, detrusor underactivity.

Article Snippet: After two freeze-thaw cycles were performed, the homogenate was centrifuged at 2°C to 8°C for 5 minutes at 5,000× g . The levels of TRPV1 and TRPV4 in the specimen were measured according to the manufacturer’s instructions using a human TRPV1 ELISA kit and a human TRPV4 ELISA kit (Cusabio Biotech Co., Houston, TX, USA).

Techniques: Immunofluorescence, Staining