Fig 1: Knockdown of TRIM29 induced cell cycle progression and promoted cell apoptosis. A and B, after down-regulating TRIM29 expression in pancreatic cancer (PC) cells, the cell cycle was arrested in the G1 phase (*p < 0.05). C western blot results showed that the expression levels of cyclins Cyclin-D1 and PCNA decreased. D apoptosis experiments confirmed that the proportion of phase apoptosis was significantly increased in shTRIM29 cells (*p < 0.05). E The expression of BAX and caspase 3 was increased in shTRIM29 cells, while that of BCL-2 was reduced. F TUNEL staining images for apoptosis in the two tumour xenograft groups. G, IHC for Ki-67 in the two tumour xenograft groups
Fig 2: TRIM29 stabilized YAP1 by regulating its ubiquitination in pancreatic cancer cells. A co-IP results showed that TRIM29 and YAP1 interacted directly. B The expression level of YAP1 increased with time in pancreatic cancer (PC) cells treated with proteasome inhibitor MG132. C and D TRIM29 did not affect YAP1 expression level, as assessed in PC cells transfected with shTRIM29/P-TRIM29 plasmid and treated with MG132. E and F Detection of YAP1 degradation using anti-HA and anti-Flag antibodies in BxPC-3 and SW1990 cells transfected with the HA-YAP1 expression plasmid, with or without the Flag-TRIM29 plasmid, and treated with cycloheximide for the indicated times. G Lysates from PC cells transduced with shTRIM29/P-TRIM29 were treated with MG132 (15 µM) for 4 h, collected, immunoprecipitated using an anti-ubiquitin (Ub) antibody, and then immunoblotted using a YAP1 antibody
Fig 3: Expression of miR-122 and TRIM29 in patients with NPC and cell lines. Quantitative analysis of (A) miR-122 and (B) TRIM29 in NPC samples with or without metastasis, respectively, was performed. p-NPC, NPC samples without metastasis; m-NPC, NPC samples with metastasis. Data are presented as the mean ± standard deviation, n=3. *P<0.05 and **P<0.01 vs. normal; #P<0.05 vs. p-NPC. (C) The overall survival rate of patients with NPC in miR-122 high and low expression level groups. (D) The overall survival rate of NPC patients in TRIM29 high and low expression level groups. Quantitative analyses of (E) miR-122 and (F) TRIM29 in NPC cell lines were performed following radiation treatment. (G) Western blot analysis and (H) determination of the expression of TRIM29 in NPC cell lines following radiation treatment. Data are presented as mean ± standard deviation, n=4. *P<0.05 and **P<0.01 vs. groups without radiation treatment. NPC, nasopharyngeal carcinoma; TRIM29, tripartite motif-containing protein 29; miR, microRNA.
Fig 4: YAP1 mediated TRIM29-induced proliferation in pancreatic cancer cells. A western blot showing TRIM29 and YAP1 expression in BxPC-3 cells stably transfected with shTRIM29 in the presence or absence of P-YAP1. B–D, EdU and CCK8 assays showed that the reduced proliferation induced by TRIM29 knockdown in BxPC-3 cells was partially abolished by the introduction of P-YAP1 (*p < 0.05). E western blot analysis of TRIM29 and YAP1 protein expression in PANC-1 cells stably transfected with P-TRIM29 in the presence or absence of shYAP1. F–H, CCK8 and EdU assays showed that the increased proliferation induced by TRIM29 overexpression in PANC-1 cells was partially abolished by the introduction of shTRIM29
Fig 5: TRIM29 positively regulated YAP1 protein level. A and B, in response to upregulating or downregulating TRIM29 expression in pancreatic cancer (PC) cells, the expression level of YAP1 protein also changed, but its mRNA level changed insignificantly (ns means not significantly). C GEPIA database analysis showed that the expression of YAP1 was significantly increased in PC (*p < 0.05, T means tumour; N means normal). D and E, Determination of YAP1 protein and mRNA levels in PC tissues and paired non-tumour tissues using western blotting and qRT-PCR. GAPDH was used as an internal control (** p < 0.01, N = Normal, T = Tumour). Representative images (G) and quantification (F) of YAP1 IHC staining in 126 paired PC and non-cancer tissues. A high YAP1 expression was observed in 62.70% (79/126) (** p < 0.01) of samples. H, Kaplan–Meier survival curves revealed that PC patients with high levels of TRIM29 expression exhibited a shorter overall and disease-free survival than those with lower levels. I Scatter plots of TRIM29 and YAP1 expression in PC tissues (*p < 0.05, r = 0.6575)
Supplier Page from Abcam for Anti-TRIM29 antibody [EPR3494]