Fig 1: TGFβ receptor kinase inhibitor blocked the effect of TGFβ on the expressions of GDF15, NDRG1 and maspin in bladder carcinoma cells. The reporter activity of SEB4 reporter vector treated with (A) various concentrations of rhTGFβ or with 20 μM SB431542, or (B) 10 ng/mL rhTGFβ treatments with (+) or without (−) 400 nM rhGDF15 or 20 μM SB431542 pretreatment, as indicated, in the HT1376 cells. The expressions of GDF15, maspin, and NDRG1 were determined using immunoblot assays (C) and quantitative analysis (D) after being treated with (+) or without (−) 10 ng/mL rhTGFβ and 20 μM SB431542 for 18 h in HT1376 cells. The quantitative data were expressed as the intensity of protein bands of the target genes/β-actin relative to the control solvent-treated group (n = 3). (E) The reporter activity of the GDF15 reporter vector was treated with various concentrations of rhTGFβ or with 20 μM SB431542 as indicated. Data are expressed as the mean percentage of luciferase activity relative to the mock-treated group (n = 6). (F) RT-4 and HT1376 cells were treated with 10 ng/mL rhTGFβ with (+) or without (−) 20 μM SB431542 pretreatment for 24 h. The supernatants of culture media (n = 4) were collected and GDF15 secretions were determined by ELISA. The data were presented as the mean percentage compared with the control group. * p < 0.05, ** p < 0.01.
Fig 2: CAPE induces the phosphorylation of ERK, JNK, and p38 to modulate the expressions of GDF15, NDRG1, and maspin in bladder carcinoma HT1376 cells. (A) HT1376 cells were treated with various concentrations of CAPE as indicated for 24 h and the mRNA levels of target genes as indicated were determined by RT-qPCR assays. (B) Protein levels of target genes, as indicated, were determined by immunoblot assays after treatment with various concentrations of CAPE as indicated for 16 h in HT1376 cells. The quantitative data were expressed as the intensity of protein bands of the target genes/β-actin relative to the control solvent-treated group (n = 3). The expressions of ERK, p-ERK (C, top), JNK, p-JNK (D, top), p38, and p-p38 (E, top) were determined by the immunoblots after 20 min of 30 μM CAPE treatment with (+) or without (−) pretreatment of the indicated MAPK inhibitors for 1 h in HT1376 cells. The protein level of GDF15, NDRG1, maspin, and β-actin of HT1376 cells after CAPE treatment with (+) or without (−) pretreatment with PD0325901 (C, bottom), SP600125 (D, bottom), or SB202190 (E, bottom). The quantitative data were expressed as the intensity of protein bands of the p-target gene/target gene or target genes/β-actin relative to the control solvent-treated group (n = 3). ∗∗P < 0.01, ∗P < 0.05.
Fig 3: Correlation between IL-17A, GDF15 and epithelial-mesenchymal transition biomarkers. (A) IL-17A and (B) GDF15 were negatively correlated with E-cadherin. (C) IL-17A was positively correlated with N-cadherin and (D) no significant correlation was observed between GDF15 and N-cadherin expression. (E) IL-17A and (F) GDF15 were positively correlated with vimentin expression. IL, interleukin; GDF, growth/differentiation factor; E-cadherin, epithelial cadherin; N-cadherin, neural cadherin.
Fig 4: GDF15 is identified as a major glycoprotein in IRPC.A Pearson correlation coefficient heatmap of quadruplicates indicates reliability of experiment. B Numbers of glycoproteins, glycopeptides, intact N-glycopeptides (IGPs), and glycans identified are presented. C Volcano plots indicate differentially expressed glycoproteins. p < 0.05. D Heatmap shows differentially expressed glycotypes. E Heatmap displays proteins modified by up-regulated glycotypes. Red indicates up-regulation, blue indicates down-regulation, and gray indicates no difference. N is HexNAc, H is Hex. F is Fucose. Cartoon symbols used for glycans (see inset) conform to the standard representation recommended by the Consortium for Functional Glycomics.
Fig 5: The boxplots of the different expression levels of the 10 ferroptosis-related genes between the normal and tumor colon tissues in the training (A) and testing (B) datasets. The heatmaps for the different expression levels of these 10 genes and the clinical characteristics between the high-risk and low-risk groups in the TCGA (C) and GSE39582 (D) datasets. Representative immunohistochemistry images (E) of FDFT1, GDF15, HAMP, and TFAP2C in CRC tissues and corresponding normal tissues. *p < 0.05, **p < 0.01, ****p < 0.0001. ns, no significance.
Supplier Page from Abcam for Anti-GDF15 antibody [EPR19939]