Journal: iScience

Article Title: SLC17A9-PTHLH-EMT axis promotes proliferation and invasion of clear renal cell carcinoma

doi: 10.1016/j.isci.2022.105764

Figure Lengend Snippet: SLC17A9 expression level was upregulated in ccRCC tissues (A) SLC17A9 mRNA expression levels in the ccRCC tissues were higher than those in the normal tissues in the TCGA cohort. (B) SLC17A9 expression in normal tissues was lower than that in paired ccRCC tissues. (C) High expression of SLC17A9 was associated with the T stage and showed an increasing trend. (D) SLC17A9 expression levels in N0 and N1 stages. (E) mRNA expression of SLC17A9 in M stage. (F) SLC17A9 expression levels in different TNM stages in TCGA. (G) SLC17A9 expression levels in G grade. (H) SLC17A9 was upregulated in tumor samples compared with normal samples in the ICGC-RECA-EU cohort. (I) SLC17A9 expression levels in tumor samples were higher than those in paired normal kidney tissues. ∗∗∗∗p < 0.0001; ∗∗∗p < 0.001; ∗∗p < 0.01; and ∗p < 0.05. SLC17A9, Solute Carrier Family 17 Member 9; ccRCC, clear cell renal cell carcinoma; KIRC, kidney renal clear cell carcinoma; TCGA, The Cancer Genome Atlas; and ICGC, International Cancer Genome Consortium. Data are represented as mean ± SD.

Article Snippet: Next, the sections were incubated with rabbit SLC17A9 primary antibody (1:100; 26731-1-AP; Proteintech, USA) at 4 °C overnight.

Techniques: Expressing

Journal: iScience

Article Title: SLC17A9-PTHLH-EMT axis promotes proliferation and invasion of clear renal cell carcinoma

doi: 10.1016/j.isci.2022.105764

Figure Lengend Snippet: Diagnostic and prognostic values of SLC17A9 (A) SLC17A9 effectively distinguished para-cancer tissues from ccRCC tissues in TCGA-KIRC (AUC = 0.9459, p < 0.0001). (B) SLC17A9 effectively distinguished para-cancer tissues from ccRCC tissues in ICGC (AUC = 0.9228, p < 0.0001). (C) Diagnostic value of SLC17A9 for ccRCC patients in TCGA-KIRC increased with time. AUC at 2 years was 0.6691, AUC at 4 years was 0.6728, and AUC at 6 years was 0.6864. (D) High SLC17A9 expression group had poorer OS than the low expression group in the TCGA cohort, p < 0.0001. (E) High SLC17A9 expression indicated poorer DFS in the TCGA cohort, p = 0.0059. (F) Survival analysis of E-MTAB-1980 validation cohort with low SLC17A9 expression group presenting better survival status, p = 0.0247. (G) Construction of a nomogram for predicting 1-, 3-, and 5-year OS possibilities of individual ccRCC patients. (H–J) Calibration curves of (H) 1-year, (I) 3-year, and (J) 5-year OS of ccRCC patients. Light-colored diagonal represented predicted results, whereas the red-colored broken line represented actual results. Data are represented as mean ± SD.

Article Snippet: Next, the sections were incubated with rabbit SLC17A9 primary antibody (1:100; 26731-1-AP; Proteintech, USA) at 4 °C overnight.

Techniques: Diagnostic Assay, Expressing, Biomarker Discovery

Journal: iScience

Article Title: SLC17A9-PTHLH-EMT axis promotes proliferation and invasion of clear renal cell carcinoma

doi: 10.1016/j.isci.2022.105764

Figure Lengend Snippet: Univariate and multivariate analyses of SLC17A9 expression and patient overall survival using TCGA-KIRC data (n = 522)

Article Snippet: Next, the sections were incubated with rabbit SLC17A9 primary antibody (1:100; 26731-1-AP; Proteintech, USA) at 4 °C overnight.

Techniques: Expressing

Journal: iScience

Article Title: SLC17A9-PTHLH-EMT axis promotes proliferation and invasion of clear renal cell carcinoma

doi: 10.1016/j.isci.2022.105764

Figure Lengend Snippet: Univariate and multivariate analyses of SLC17A9 expression and patient disease free survival. using TCGA-KIRC data (n = 522)

Article Snippet: Next, the sections were incubated with rabbit SLC17A9 primary antibody (1:100; 26731-1-AP; Proteintech, USA) at 4 °C overnight.

Techniques: Expressing

Journal: iScience

Article Title: SLC17A9-PTHLH-EMT axis promotes proliferation and invasion of clear renal cell carcinoma

doi: 10.1016/j.isci.2022.105764

Figure Lengend Snippet: SLC17A9 was upregulated in ccRCC cell lines and tissues (A and B) Relative mRNA and protein levels of SLC17A9 in ccRCC cell lines. (C and D) Relative mRNA and protein levels of SLC17A9 in ccRCC tissues and adjacent normal kidney tissues. (E) Immunohistochemistry (IHC) results (200x,400x) of SLC17A9 in ccRCC tissues and para-cancer tissues. ∗∗∗∗p < 0.0001; ∗∗∗p < 0.001; ∗∗p < 0.01; and ∗p < 0.05. Data are represented as mean ± SD. The length of scale bars are 50 and 20 μm.

Article Snippet: Next, the sections were incubated with rabbit SLC17A9 primary antibody (1:100; 26731-1-AP; Proteintech, USA) at 4 °C overnight.

Techniques: Immunohistochemistry

Journal: iScience

Article Title: SLC17A9-PTHLH-EMT axis promotes proliferation and invasion of clear renal cell carcinoma

doi: 10.1016/j.isci.2022.105764

Figure Lengend Snippet: Effects of SLC17A9 on the proliferation of renal cancer cells (A and B) Successful knockdown of SLC17A9 mRNA in 786-O and A498 cells. (C and D) SLC17A9 mRNA levels were significantly overexpressed in 786-O and A498 cells. (E and F) Protein levels after SLC17A9 knockdown in 786-O and A498 cells. (G and H) Protein levels after SLC17A9 overexpression in 786-O and A498 cells. (I–L) Cell counting kit-8 assays detected the effects of SLC17A9 overexpression and knockdown on the proliferation of 786-O and A498 cells. ∗∗∗∗p < 0.0001; ∗∗∗p < 0.001; ∗∗p < 0.01; and ∗p < 0.05. Data were presented as the mean ± standard deviation (SD) from three independent experiments.

Article Snippet: Next, the sections were incubated with rabbit SLC17A9 primary antibody (1:100; 26731-1-AP; Proteintech, USA) at 4 °C overnight.

Techniques: Knockdown, Over Expression, Cell Counting, Standard Deviation

Journal: iScience

Article Title: SLC17A9-PTHLH-EMT axis promotes proliferation and invasion of clear renal cell carcinoma

doi: 10.1016/j.isci.2022.105764

Figure Lengend Snippet: SLC17A9 affected renal cancer cells via EMT signaling pathways (A) GSEA results of SLC17A9 using TCGA data. (B) Correlation analyses of Snail, N-cadherin, E-cadherin, and vimentin with SLC17A9 were performed using the TCGA-KIRC data. (C–F) Effects of SLC17A9 overexpression and knockdown on E-cadherin, N-cadherin, vimentin, and snail-1 in renal cancer cells. (G–J) Effects of SLC17A9 overexpression and knockdown on cell migration and invasion. ∗∗∗∗p < 0.0001. Data are represented as mean ± SD. The length of scale bar is 100 μm.

Article Snippet: Next, the sections were incubated with rabbit SLC17A9 primary antibody (1:100; 26731-1-AP; Proteintech, USA) at 4 °C overnight.

Techniques: Protein-Protein interactions, Over Expression, Knockdown, Migration

Journal: iScience

Article Title: SLC17A9-PTHLH-EMT axis promotes proliferation and invasion of clear renal cell carcinoma

doi: 10.1016/j.isci.2022.105764

Figure Lengend Snippet: SLC17A9-activated EMT pathway was dependent on PTHLH (A) Four potential downstream genes of SLC17A9. (B and C) Regulation of PTHLH by SLC17A9 was verified at the mRNA level. (D) Regulation of PTHLH by SLC17A9 was verified at the protein level. (E and F) Knockdown of PTHLH could reverse the effect of SLC17A9 overexpression on EMT markers. (G and H) Migration and invasion assays (200x) for the indicated ccRCC cells. ∗∗∗∗p < 0.0001; ∗∗∗p < 0.001; ∗∗p < 0.01; and ∗p < 0.05. Data are represented as mean ± SD. The length of scale bar is 100 μm.

Article Snippet: Next, the sections were incubated with rabbit SLC17A9 primary antibody (1:100; 26731-1-AP; Proteintech, USA) at 4 °C overnight.

Techniques: Knockdown, Over Expression, Migration

Journal: iScience

Article Title: SLC17A9-PTHLH-EMT axis promotes proliferation and invasion of clear renal cell carcinoma

doi: 10.1016/j.isci.2022.105764

Figure Lengend Snippet: Upregulation of SLC17A9 promoted tumor progression in vivo (A) Prediction of SLC17A9 associated drugs. (B and C) IC 50 of vorinostat in A498 and 786-O. (D) Protein expression levels of SLC17A9 in cells cultivated at different vorinostat concentrations. (E–I) Subcutaneous tumor in mice, its growth curve, and final volume and weight (n = 5). ∗∗∗∗p < 0.0001; ∗∗∗p < 0.001; ∗∗p < 0.01; and ∗p < 0.05. Data are represented as mean ± SD.

Article Snippet: Next, the sections were incubated with rabbit SLC17A9 primary antibody (1:100; 26731-1-AP; Proteintech, USA) at 4 °C overnight.

Techniques: In Vivo, Expressing

Journal: iScience

Article Title: SLC17A9-PTHLH-EMT axis promotes proliferation and invasion of clear renal cell carcinoma

doi: 10.1016/j.isci.2022.105764

Figure Lengend Snippet:

Article Snippet: Next, the sections were incubated with rabbit SLC17A9 primary antibody (1:100; 26731-1-AP; Proteintech, USA) at 4 °C overnight.

Techniques: Recombinant, Software

Journal: Discover. Oncology

Article Title: Pyroptosis-related genes regulate proliferation and invasion of pancreatic cancer and serve as the prognostic signature for modeling patient survival.

doi: 10.1007/s12672-022-00495-0

Figure Lengend Snippet: Fig. 5 Expression and survival analysis of PRGs in PDAC. A The protein expression score of staining and intensity of NLRP1 and CASP4 pro- teins in all PDAC samples (HPA database). Protein expression score is based on immunohistochemical data manually scored with regard to staining intensity (negative, weak, moderate or strong) and fraction of stained cells (< 25%, 25–75% or > 75%). B Immunohistochemi- cal staining of NLRP1 and CASP4 proteins in PDAC. C Methylation levels of CASP4 at chr11:104838825–104841325 and chr11:104826922– 104829422 in PDAC and para-cancer tissues. D The OS survival curves of NLRP1 and CASP4 mRNA expression level in PDAC. E CASP4 and NLRP1 immunohistochemistry for patients from Zhongnan Hospital (×200)

Article Snippet: To test whether the expression of CASP4 or NLRP1 affects the sensitivity of pancreatic cancer cells to MEK inhibitors, Trametinib (MedChemExpress, USA) was added to siRNA-transfected PANC-1 and Aspc-1 cells.

Techniques: Expressing, Staining, Immunohistochemical staining, Methylation, Immunohistochemistry

Journal: Discover. Oncology

Article Title: Pyroptosis-related genes regulate proliferation and invasion of pancreatic cancer and serve as the prognostic signature for modeling patient survival.

doi: 10.1007/s12672-022-00495-0

Figure Lengend Snippet: Fig. 6 PRGs regulated the proliferation and invasion of pancreatic cancer cells in vitro. A, B After siRNA transfection, qPCR and western blot was used to detect RNA and protein expression levels of CASP4 and NLRP1 respectively in PANC-1 and Aspc-1 cells. C CCK8 assay was used to detect the proliferation of PANC-1 and Aspc-1 cells after transfection with siRNA. D Transwell was used to detect the change of cell inva- sion ability after transfection with siRNA (*P < 0.05, **P < 0.01, ***P < 0.001)

Article Snippet: To test whether the expression of CASP4 or NLRP1 affects the sensitivity of pancreatic cancer cells to MEK inhibitors, Trametinib (MedChemExpress, USA) was added to siRNA-transfected PANC-1 and Aspc-1 cells.

Techniques: In Vitro, Transfection, Western Blot, Expressing, CCK-8 Assay

Journal: Discover. Oncology

Article Title: Pyroptosis-related genes regulate proliferation and invasion of pancreatic cancer and serve as the prognostic signature for modeling patient survival.

doi: 10.1007/s12672-022-00495-0

Figure Lengend Snippet: Fig. 9 NLRP1 was associated with KRAS and P53 mutations potentially. A The expression levels of NLRP1 were different in KRAS mutation, P53 mutation and wild-type tumor tissues respectively. B Correlation curve between NLRP1, KRAS and P53 expression levels based on TCGA database. C GSEA analysis of DEGs in high NLRP1 expression group. D PANC-1 and Aspc-1 cells were tested for LDH release 24 h after siNLRP1 transfection. E Cleavage of GSDMD and p-ERK were monitored by immunoblot analysis 48 h after siNLRP1 transfection (*P < 0.05, ***P < 0.001)

Article Snippet: To test whether the expression of CASP4 or NLRP1 affects the sensitivity of pancreatic cancer cells to MEK inhibitors, Trametinib (MedChemExpress, USA) was added to siRNA-transfected PANC-1 and Aspc-1 cells.

Techniques: Expressing, Mutagenesis, Transfection, Western Blot

Journal: Discover. Oncology

Article Title: Pyroptosis-related genes regulate proliferation and invasion of pancreatic cancer and serve as the prognostic signature for modeling patient survival.

doi: 10.1007/s12672-022-00495-0

Figure Lengend Snippet: Fig. 10 Analysis of drug sensitivity associated with PEGs. A Correlation of CASP4 and NLRP1 expression with multidrug sensitivity in GDSC. Red is positive correlation, which means the higher the gene expression, the more sensitive to the drug, while the blue is the opposite. The Spearman correlation represent the gene expression correlates with the drug. The positive correlation means that the gene high expression is resistant to the drug, vise verse. B Survival rate of cells exposed to trametinib at the concentrations of 10 µM/L for 48 or 72 h (*P < 0.05, **P < 0.01, ***P < 0.001)

Article Snippet: To test whether the expression of CASP4 or NLRP1 affects the sensitivity of pancreatic cancer cells to MEK inhibitors, Trametinib (MedChemExpress, USA) was added to siRNA-transfected PANC-1 and Aspc-1 cells.

Techniques: Expressing, Gene Expression

Journal: Chemical Engineering Journal

Article Title: Self-healing hydrogel reduces inflammation through ANT1/OPTN axis mediated mitophagy for spinal cord injury repair

doi: 10.1016/j.cej.2024.152263

Figure Lengend Snippet: Fig. 6. RNA-seq analysis revealed that ANT1-mediated mitophagy involves in anti-inflammation effects. The top 10 upregulated enriched GO term (a) and the top 10 downregulated enriched GO term (b) of differentially expressed (p < 0.05, |log2FC > 0.58|) mRNA in TPHD@Tc treated zebrafish compared with SCI group. (c) Veen analysis of the differentially expressed mRNA and autophagy-related genes. mRNA expression (d, n = 3), protein expression (e, n = 3) and Immunoflu orescent staining (f) of ANT1 in BV2 cells treated with TPHD, TPHD@TcL, TPHD@Tc and TPHD@TcH for 4 days. mRNA (g, n = 3), protein expression (h, n = 3) and Immunofluorescent staining (i) of NLRP3 Data were expressed by mean with SD. * p < 0.05, ** p < 0.01, **** p < 0.0001, compared with TPHD in b and c.

Article Snippet: The primary antibodies of NLRP3 (1:1000, Rabbit, ab263899, Abcam), ANT1 (1:1000, Rabbit, NBP292642, NOVUS), and the secondary antibodies of HRP-conjugated Affinipure Goat Anti-Rabbit IgG (H + L) (1:5000, SA00001-2, Proteintech) were used.

Techniques: RNA Sequencing, Expressing, Staining

Journal: Chemical Engineering Journal

Article Title: Self-healing hydrogel reduces inflammation through ANT1/OPTN axis mediated mitophagy for spinal cord injury repair

doi: 10.1016/j.cej.2024.152263

Figure Lengend Snippet: Fig. 7. ANT1 enhanced OPTN recruitment promotes mitophagy mediated inflammation inhibition. Immunofluorescence of TOMM20 (red) and OPTN (green) in BV2 cells treated with TPHD (a), TPHD@TcL (b), TPHD@Tc (c), TPHD@TcH (d) for 4 days. LC3B expression in BV2 cells treated with TPHD, TPHD@TcL, TPHD@Tc, TPHD@TcH for 4 days (e) and related quantitative m-cherry labeled area (f, n = 5), GFP labeled area (g, n = 5) and m-cherry and GFP co-labeled area (h, n = 5). NLRP3 mRNA expression (i, n = 3) and protein expression (j, n = 3) in BV2 cells treated with TPHD, TPHD@TcL, TPHD@Tc, TPHD@TcH for 4 days. Immunofluorescent staining of NLRP3 of BV2 cells treated with TPHD, TPHD@TcL, TPHD@Tc, TPHD@TcH for 4 days (k). (l) The schematic diagram of hydrogel containing Tc promoted mitophagy to decrease inflammation. The elevated ANT1 promoted OPTN recruitment, which provided LC3 anchoring for phagophore formation around dysfunctional mitochondria, leading to a regressive inflammation. Data were expressed by mean with SD. * p < 0.05, *** p < 0.001, **** p < 0.0001, compared with TPHD in f-j.

Article Snippet: The primary antibodies of NLRP3 (1:1000, Rabbit, ab263899, Abcam), ANT1 (1:1000, Rabbit, NBP292642, NOVUS), and the secondary antibodies of HRP-conjugated Affinipure Goat Anti-Rabbit IgG (H + L) (1:5000, SA00001-2, Proteintech) were used.

Techniques: Inhibition, Immunofluorescence, Expressing, Labeling, Staining