antibodies against trpa1  (Novus Biologicals)

Journal: Molecular Oncology

Article Title: TRPM8 levels determine tumor vulnerability to channel agonists

doi: 10.1002/1878-0261.70049

Figure Lengend Snippet: TRPM8 RNA and protein expression in solid cancers. (A) Landscape of TRPM8 transcript levels in primary tumor samples across different tissues (horizontal axis) was retrieved from The Cancer Genome Atlas (TCGA) datasets of cBioPortal. Panel display boxplots illustrating data distributions; boxes represent the median and interquartile range, while whiskers indicate variability beyond the quartiles, with individual data points shown above the plots. (B) Representative images of TRPM8 immunolocalization in healthy and malignant prostate, colorectal, breast, and lung tissue samples spotted on a commercial tissue microarray (TMA). TRPM8 immunostaining was scored as absent (0), weak (1), moderate (2) or high (3) (scale bar 20 μm). (C) TRPM8 score related to tumor stage ( n = 48 cores per cancer type; n = 4 cores per normal tissue type). (D) Direct comparison of TRPM8 RNA expression across prostate, colorectal, breast, and lung cancers. Panel display boxplots illustrating data distributions; boxes represent the median and interquartile range, while whiskers indicate variability beyond the quartiles, with individual data points shown above the plots. (E, F) Quantification of TRPM8 RNA expression and protein amount in matched normal prostate tissue (N) and prostate cancer (T) samples isolated from n = 3 radical prostatectomies of prostate cancer (PCa) patients (E), and n = 5 independent colorectal cancer (CRC) samples (F). (G) Western blot of TRPM8 protein in the samples described in (E, F).

Article Snippet: To analyze the status of the TRPM8 channel in a group of solid tumors other than prostate cancer, a Tissue Microarray was purchased from US Biomax, Inc. (MC2081a) and stained with a validated antibody against TRPM8 (Alomone #ACC‐049; [ , , ]) at the Anatomic Pathology Operative Unit of the Santa Chiara Hospital of Trento.

Techniques: Expressing, Microarray, Immunostaining, Comparison, RNA Expression, Isolation, Western Blot

Journal: Molecular Oncology

Article Title: TRPM8 levels determine tumor vulnerability to channel agonists

doi: 10.1002/1878-0261.70049

Figure Lengend Snippet: Variable amounts of TRPM8 channel in tumor cells with low levels of its coding transcript. (A) TRPM8 RNA expression in cancer cell lines described in the NCI‐60 cell lines and Cancer Cell Lines Encyclopedia projects (retrieved from cBioPortal). (B) Quantitative reverse transcription polymerase chain reaction (RT‐qPCR) comparative analysis of TRPM8 RNA expression in prostate (VCaP, LNCaP, PC3), colon (HCT116), breast (MCF7) and lung (A549) cancer cells. (C–F) Western blot replicas I and II of TRPM8 in VCaP, LNCaP, PC3, HCT116, MCF7, A549 cell lines with the Alomone ACC‐049 (C) and Abcam Ab3243 (E) antibodies, and relative quantification of TRPM8 protein (D, F) in the n = 4 independent replicas shown in C–E and Fig. . β‐Actin is used as loading control and normalizer. Data are presented as mean ± standard deviation (sd) of three (Alomone ACC‐049 antibody) and four (Abcam Ab3243 antibody) independent experiments. *** P ≤ 0.001, ** P ≤ 0.01, * P ≤ 0.05. Statistical analysis was performed using Student's t ‐test.

Article Snippet: To analyze the status of the TRPM8 channel in a group of solid tumors other than prostate cancer, a Tissue Microarray was purchased from US Biomax, Inc. (MC2081a) and stained with a validated antibody against TRPM8 (Alomone #ACC‐049; [ , , ]) at the Anatomic Pathology Operative Unit of the Santa Chiara Hospital of Trento.

Techniques: RNA Expression, Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Quantitative Proteomics, Control, Standard Deviation

Journal: Molecular Oncology

Article Title: TRPM8 levels determine tumor vulnerability to channel agonists

doi: 10.1002/1878-0261.70049

Figure Lengend Snippet: Lethal synergy between TRPM8 agonist D‐3263 and chemotherapy in cancer cells. (A) Representative crystal violet staining of LNCaP, HCT116, MCF7, A549, and PC3 cells untreated or treated with the indicated drugs for 24 h (chemotherapy = Docetaxel for LNCaP, MCF7, A549, and PC3; 5‐fluorouracil (5‐FU + Oxaliplatin for HCT116). TRPM8 knock‐down (siRNA1 and siRNA2) confirmed D‐3263 specificity. (B) Quantification of the viability of treated cells compared with untreated controls. Data are presented as mean ± standard deviation (sd) of n = 3 independent experiments (A, Fig. ). *** P < 0.001. Statistical analysis was performed using Student's t ‐test. (C, D) Western blot analysis of Caspase 3 and Parp cleavage in LNCaP, HCT116, MCF7, A549, and PC3 cells untreated or treated with the indicated drugs for 24 h (C). TRPM8 knock‐down (siRNA2) confirmed D‐3263 specificity (D). (E) Direct comparison of D‐3263 efficacy in wild type and TRPM8 knocked down (KD) cancer cells. Western blot analysis in C–E was repeated with n = 2 sets of biologically independent samples.

Article Snippet: To analyze the status of the TRPM8 channel in a group of solid tumors other than prostate cancer, a Tissue Microarray was purchased from US Biomax, Inc. (MC2081a) and stained with a validated antibody against TRPM8 (Alomone #ACC‐049; [ , , ]) at the Anatomic Pathology Operative Unit of the Santa Chiara Hospital of Trento.

Techniques: Staining, Knockdown, Standard Deviation, Western Blot, Comparison

Journal: Molecular Oncology

Article Title: TRPM8 levels determine tumor vulnerability to channel agonists

doi: 10.1002/1878-0261.70049

Figure Lengend Snippet: Efficacy of D‐3263 + 5‐FU + Oxaliplatin in CRC organoids raises with the amount of TRPM8. (A) TRPM8 immunolocalization in a dedicated homemade tissue microarray (TMA) of colorectal cancer (scale bar 100 μm; ACC‐049). Representative images of colorectal cancer (CRC) with different levels of TRPM8 staining and relative scores. Score 1: weak expression, score 2: moderate expression, score 3: high expression. (B) Distribution of TRPM8 immunostaining scores across the colorectal cancer (CRC) specimens ( n = 79 independent cores). (C) Immunolocalization of TRPM8 in n = 5 different lines of patient‐derived colorectal cancer (CRC) organoids (scale bar 20 μm). (D) Western blot of TRPM8 from colorectal cancer (CRC) specimens ( n = 5 independent samples) derived from the tissue bank of the Santa Chiara Hospital of Trento and—independent— n = 5 patient‐derived organoids (PDO). Western blot analysis in D was repeated twice. (E) Relative viability of colorectal cancer (CRC) organoids subjected to the indicated treatments or left untreated to serve as control pooled together based on TRPM8 protein amount (high (Hi): PDO #1 and PDO #4; low (Lo): PDO #2, PDO #3 and PDO #5). Distribution of viability measures across the different conditions and stratifications is shown using boxplots. Paired two‐sample t ‐test was used to compare the viability of organoid lines. Data are presented as mean ± standard deviation (sd) of two independent biological experiments, each with six technical replicates.

Article Snippet: To analyze the status of the TRPM8 channel in a group of solid tumors other than prostate cancer, a Tissue Microarray was purchased from US Biomax, Inc. (MC2081a) and stained with a validated antibody against TRPM8 (Alomone #ACC‐049; [ , , ]) at the Anatomic Pathology Operative Unit of the Santa Chiara Hospital of Trento.

Techniques: Microarray, Staining, Expressing, Immunostaining, Derivative Assay, Western Blot, Control, Standard Deviation

Journal: Molecular Oncology

Article Title: TRPM8 levels determine tumor vulnerability to channel agonists

doi: 10.1002/1878-0261.70049

Figure Lengend Snippet: TRPM8 RNA and protein expression in solid cancers. (A) Landscape of TRPM8 transcript levels in primary tumor samples across different tissues (horizontal axis) was retrieved from The Cancer Genome Atlas (TCGA) datasets of cBioPortal. Panel display boxplots illustrating data distributions; boxes represent the median and interquartile range, while whiskers indicate variability beyond the quartiles, with individual data points shown above the plots. (B) Representative images of TRPM8 immunolocalization in healthy and malignant prostate, colorectal, breast, and lung tissue samples spotted on a commercial tissue microarray (TMA). TRPM8 immunostaining was scored as absent (0), weak (1), moderate (2) or high (3) (scale bar 20 μm). (C) TRPM8 score related to tumor stage ( n = 48 cores per cancer type; n = 4 cores per normal tissue type). (D) Direct comparison of TRPM8 RNA expression across prostate, colorectal, breast, and lung cancers. Panel display boxplots illustrating data distributions; boxes represent the median and interquartile range, while whiskers indicate variability beyond the quartiles, with individual data points shown above the plots. (E, F) Quantification of TRPM8 RNA expression and protein amount in matched normal prostate tissue (N) and prostate cancer (T) samples isolated from n = 3 radical prostatectomies of prostate cancer (PCa) patients (E), and n = 5 independent colorectal cancer (CRC) samples (F). (G) Western blot of TRPM8 protein in the samples described in (E, F).

Article Snippet: TMA sections were stained with two independent specific antibodies against TRPM8 (Alomone ACC‐049 and Abcam Ab3243) and analyzed by three experienced pathologists (MB, FGC, GB).

Techniques: Expressing, Microarray, Immunostaining, Comparison, RNA Expression, Isolation, Western Blot

Journal: Molecular Oncology

Article Title: TRPM8 levels determine tumor vulnerability to channel agonists

doi: 10.1002/1878-0261.70049

Figure Lengend Snippet: Variable amounts of TRPM8 channel in tumor cells with low levels of its coding transcript. (A) TRPM8 RNA expression in cancer cell lines described in the NCI‐60 cell lines and Cancer Cell Lines Encyclopedia projects (retrieved from cBioPortal). (B) Quantitative reverse transcription polymerase chain reaction (RT‐qPCR) comparative analysis of TRPM8 RNA expression in prostate (VCaP, LNCaP, PC3), colon (HCT116), breast (MCF7) and lung (A549) cancer cells. (C–F) Western blot replicas I and II of TRPM8 in VCaP, LNCaP, PC3, HCT116, MCF7, A549 cell lines with the Alomone ACC‐049 (C) and Abcam Ab3243 (E) antibodies, and relative quantification of TRPM8 protein (D, F) in the n = 4 independent replicas shown in C–E and Fig. . β‐Actin is used as loading control and normalizer. Data are presented as mean ± standard deviation (sd) of three (Alomone ACC‐049 antibody) and four (Abcam Ab3243 antibody) independent experiments. *** P ≤ 0.001, ** P ≤ 0.01, * P ≤ 0.05. Statistical analysis was performed using Student's t ‐test.

Article Snippet: TMA sections were stained with two independent specific antibodies against TRPM8 (Alomone ACC‐049 and Abcam Ab3243) and analyzed by three experienced pathologists (MB, FGC, GB).

Techniques: RNA Expression, Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Quantitative Proteomics, Control, Standard Deviation

Journal: Molecular Oncology

Article Title: TRPM8 levels determine tumor vulnerability to channel agonists

doi: 10.1002/1878-0261.70049

Figure Lengend Snippet: Lethal synergy between TRPM8 agonist D‐3263 and chemotherapy in cancer cells. (A) Representative crystal violet staining of LNCaP, HCT116, MCF7, A549, and PC3 cells untreated or treated with the indicated drugs for 24 h (chemotherapy = Docetaxel for LNCaP, MCF7, A549, and PC3; 5‐fluorouracil (5‐FU + Oxaliplatin for HCT116). TRPM8 knock‐down (siRNA1 and siRNA2) confirmed D‐3263 specificity. (B) Quantification of the viability of treated cells compared with untreated controls. Data are presented as mean ± standard deviation (sd) of n = 3 independent experiments (A, Fig. ). *** P < 0.001. Statistical analysis was performed using Student's t ‐test. (C, D) Western blot analysis of Caspase 3 and Parp cleavage in LNCaP, HCT116, MCF7, A549, and PC3 cells untreated or treated with the indicated drugs for 24 h (C). TRPM8 knock‐down (siRNA2) confirmed D‐3263 specificity (D). (E) Direct comparison of D‐3263 efficacy in wild type and TRPM8 knocked down (KD) cancer cells. Western blot analysis in C–E was repeated with n = 2 sets of biologically independent samples.

Article Snippet: TMA sections were stained with two independent specific antibodies against TRPM8 (Alomone ACC‐049 and Abcam Ab3243) and analyzed by three experienced pathologists (MB, FGC, GB).

Techniques: Staining, Knockdown, Standard Deviation, Western Blot, Comparison

Journal: Molecular Oncology

Article Title: TRPM8 levels determine tumor vulnerability to channel agonists

doi: 10.1002/1878-0261.70049

Figure Lengend Snippet: Efficacy of D‐3263 + 5‐FU + Oxaliplatin in CRC organoids raises with the amount of TRPM8. (A) TRPM8 immunolocalization in a dedicated homemade tissue microarray (TMA) of colorectal cancer (scale bar 100 μm; ACC‐049). Representative images of colorectal cancer (CRC) with different levels of TRPM8 staining and relative scores. Score 1: weak expression, score 2: moderate expression, score 3: high expression. (B) Distribution of TRPM8 immunostaining scores across the colorectal cancer (CRC) specimens ( n = 79 independent cores). (C) Immunolocalization of TRPM8 in n = 5 different lines of patient‐derived colorectal cancer (CRC) organoids (scale bar 20 μm). (D) Western blot of TRPM8 from colorectal cancer (CRC) specimens ( n = 5 independent samples) derived from the tissue bank of the Santa Chiara Hospital of Trento and—independent— n = 5 patient‐derived organoids (PDO). Western blot analysis in D was repeated twice. (E) Relative viability of colorectal cancer (CRC) organoids subjected to the indicated treatments or left untreated to serve as control pooled together based on TRPM8 protein amount (high (Hi): PDO #1 and PDO #4; low (Lo): PDO #2, PDO #3 and PDO #5). Distribution of viability measures across the different conditions and stratifications is shown using boxplots. Paired two‐sample t ‐test was used to compare the viability of organoid lines. Data are presented as mean ± standard deviation (sd) of two independent biological experiments, each with six technical replicates.

Article Snippet: TMA sections were stained with two independent specific antibodies against TRPM8 (Alomone ACC‐049 and Abcam Ab3243) and analyzed by three experienced pathologists (MB, FGC, GB).

Techniques: Microarray, Staining, Expressing, Immunostaining, Derivative Assay, Western Blot, Control, Standard Deviation

Journal: Cancer medicine

Article Title: TRPM8 overexpression suppresses hepatocellular carcinoma progression and improves survival by modulating the RTP3/STAT3 pathway.

doi: 10.1002/cam4.70109

Figure Lengend Snippet: FIGURE 4 TRPM8 level determination and TRPM8-silencing cell function experiments. (A–D) TRPM8 inhibition in PLC/PRF/5 and HUH-7 cells transfected with shTRPM8 lentivirus was verified by Western blot and qRT-PCR. (E–I) Effect of TRPM8 loss-of-function on PLC/RPF/5 and HUH-7 cell proliferation was determined by colony formation assay (E), CCK-8 (F-G) and EdU assay (H, I). (J, K) The effects of TRPM8 inhibition on the cell migration and invasion of PLC/PRF/5 and HUH-7 cells were detected using Transwell assays. Scale bars: 100 μm. *p < 0.05.

Article Snippet: Primary antibodies against TRPM8 (Zen- bioscience, Chengdu, China), RTP3 (Epigentek, USA), STAT3 (Proteintech, Wuhan, China) and GAPDH (Proteintech, Wuhan, China) were employed.

Techniques: Cell Function Assay, Inhibition, Transfection, Western Blot, Quantitative RT-PCR, Colony Assay, CCK-8 Assay, EdU Assay, Migration

Journal: Cancer medicine

Article Title: TRPM8 overexpression suppresses hepatocellular carcinoma progression and improves survival by modulating the RTP3/STAT3 pathway.

doi: 10.1002/cam4.70109

Figure Lengend Snippet: FIGURE 6 AD80 promoted the expression of TRPM8 and inhibited the proliferation, migration and invasion of HCC cells in vitro. (A, B) Calcium imaging was used to detected the changes of the fluorescence intensity of WS12 (A) and AD80 (B) groups on time in SNU-449 cells. (C, D) Effects of WS12 (C) and AD80 (D) on the expression of TRPM8 in SNU-449 cells were detected by Western blot. (E-G) EdU (E) and CCK-8 assays (F) were used to detect the effect of WS12 on the proliferation of SNU-449. (G) The effects of WS12 on the cell migration and invasion of SSNU-449 cells were detected using Transwell assays. (H–J) EdU (H) and CCK-8 assays (I) were used to detect the effect of WS12 on the proliferation of SSNU-449. (J) The effects of AD80 on the cell migration and invasion of SNU-449 cells were detected using Transwell assays. Scale bars: 100 μm. *p < 0.05.

Article Snippet: Primary antibodies against TRPM8 (Zen- bioscience, Chengdu, China), RTP3 (Epigentek, USA), STAT3 (Proteintech, Wuhan, China) and GAPDH (Proteintech, Wuhan, China) were employed.

Techniques: Expressing, Migration, In Vitro, Imaging, Fluorescence, Western Blot, CCK-8 Assay

Journal: Cancer medicine

Article Title: TRPM8 overexpression suppresses hepatocellular carcinoma progression and improves survival by modulating the RTP3/STAT3 pathway.

doi: 10.1002/cam4.70109

Figure Lengend Snippet: FIGURE 7 Schematic depicting the possible mechanism of the TRPM8- mediated RTP3/STAT3 pathway in HCC.

Article Snippet: Primary antibodies against TRPM8 (Zen- bioscience, Chengdu, China), RTP3 (Epigentek, USA), STAT3 (Proteintech, Wuhan, China) and GAPDH (Proteintech, Wuhan, China) were employed.

Techniques: