rabbit anti fam83a  (Proteintech)

Journal: Journal of Oncology

Article Title: Overexpression of FAM83A Is Associated with Poor Prognosis of Lung Adenocarcinoma

doi: 10.1155/2022/8767333

Figure Lengend Snippet: Correlations between FAM83A expression and clinical/pathological characteristics in LUAD.

Article Snippet: The tissue pieces were washed and covered with rabbit anti-FAM83A antibody (1 : 400; No: orb183622, Biorbyt, Cambridge, UK) and incubated overnight at 4°C.

Techniques: Expressing

Journal: Journal of Oncology

Article Title: Overexpression of FAM83A Is Associated with Poor Prognosis of Lung Adenocarcinoma

doi: 10.1155/2022/8767333

Figure Lengend Snippet: UALCAN analysis of TCGA showed FAM83A is overexpressed in LUAD. (a) UALCAN analysis showed FAM83A is overexpressed in lung adenocarcinoma (LUAD) and lung squamous carcinoma (LUSC), especially the former. (b, c) Cluster analysis showed a significant over expression of FAM83A in LUAD (tumor) than in normal lung tissue (normal). (d) Kaplan-Meier analysis showed that patients with high FAM83A expression had shorter overall survival, P < 0.0001.

Article Snippet: The tissue pieces were washed and covered with rabbit anti-FAM83A antibody (1 : 400; No: orb183622, Biorbyt, Cambridge, UK) and incubated overnight at 4°C.

Techniques: Over Expression, Expressing

Journal: Journal of Oncology

Article Title: Overexpression of FAM83A Is Associated with Poor Prognosis of Lung Adenocarcinoma

doi: 10.1155/2022/8767333

Figure Lengend Snippet: FAM83A protein was overexpressed in LUAD and higher FAM83A expressions related to poor overall patient survival. (a) Representative hematein-eosin (HE) and immunohistochemistry (IHC) images of FAM83A expressions in adjacent normal tissues (NORMAL) and LUAD with low (LOW) and high (HIGH) FAM83A expressions. The short bar is equal to 50 microns; the long bar is equal to 100 microns. (b) The average optical density (AOD) of FAM83A in LUAD tissues and adjacent normal lung tissues. (c) AOD of FAM83A in LUAD tissues in stage I-II and stage III-IV. (d) Kaplan-Meier survival analysis showed a significant difference in 84 LUAD patients grouped by low and high FAM83A expression. ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: The tissue pieces were washed and covered with rabbit anti-FAM83A antibody (1 : 400; No: orb183622, Biorbyt, Cambridge, UK) and incubated overnight at 4°C.

Techniques: Immunohistochemistry, Expressing

Journal: Journal of Oncology

Article Title: Overexpression of FAM83A Is Associated with Poor Prognosis of Lung Adenocarcinoma

doi: 10.1155/2022/8767333

Figure Lengend Snippet: Univariate and multivariate analyses for over survival in patients with LUAD.

Article Snippet: The tissue pieces were washed and covered with rabbit anti-FAM83A antibody (1 : 400; No: orb183622, Biorbyt, Cambridge, UK) and incubated overnight at 4°C.

Techniques:

Journal: Journal of Oncology

Article Title: Overexpression of FAM83A Is Associated with Poor Prognosis of Lung Adenocarcinoma

doi: 10.1155/2022/8767333

Figure Lengend Snippet: FAM83Apromoted proliferation of LUAD cells. (a, b) Western blotting analysis showed that FAM83A levels were higher in A549 and H1795 cells. (c) The reverse transcription-polymerase chain reaction (RT-PCR) assay demonstrated that A549 and H1795 cells transfected with FAM83A lentivirus (A549/H1795-shFAM83A), showed less FAM83A RNA expressions. (d) The cell proliferation assay showed no differences between A549-shFAM83A and A549-shRNA-NC cells at each time point. (e) The cell proliferation assay showed significant differences between H1795-shFAM83A and H1795-shRNA-NC cells. ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001.

Article Snippet: The tissue pieces were washed and covered with rabbit anti-FAM83A antibody (1 : 400; No: orb183622, Biorbyt, Cambridge, UK) and incubated overnight at 4°C.

Techniques: Western Blot, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Transfection, Proliferation Assay, shRNA

Journal: Journal of Oncology

Article Title: Overexpression of FAM83A Is Associated with Poor Prognosis of Lung Adenocarcinoma

doi: 10.1155/2022/8767333

Figure Lengend Snippet: FAM83A promoted LUAD cell migration ability. (a, b) The wound-healing assay showed significantly larger healing area in A549-shFAM83A and H1795-shFAM83A cells. (c, d) The transwell cell migration assay showed that in A549-shFAM83A and H1795-shFAM83A, more cells passed through the basement membrane. ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: The tissue pieces were washed and covered with rabbit anti-FAM83A antibody (1 : 400; No: orb183622, Biorbyt, Cambridge, UK) and incubated overnight at 4°C.

Techniques: Migration, Wound Healing Assay, Cell Migration Assay, Membrane

Journal: Journal of Oncology

Article Title: Overexpression of FAM83A Is Associated with Poor Prognosis of Lung Adenocarcinoma

doi: 10.1155/2022/8767333

Figure Lengend Snippet: FAM83A induced EMT in LUAD cells. (a, b) Western blotting showed that FAM83A and E-cadherin expression increased, while Vimentin and Snail decreased when FAM83A was knocked down in A549 cells. (c, d) The same results were seen in H1795 cells. β -Actin was used as the loading control. ∗∗∗ P < 0.001.

Article Snippet: The tissue pieces were washed and covered with rabbit anti-FAM83A antibody (1 : 400; No: orb183622, Biorbyt, Cambridge, UK) and incubated overnight at 4°C.

Techniques: Western Blot, Expressing, Control

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Positive feedback loop of FAM83A/PI3K/AKT/c-Jun induces migration, invasion and metastasis in hepatocellular carcinoma.

doi: 10.1016/j.biopha.2019.109780

Figure Lengend Snippet: Fig. 1. FAM83A was overexpressed in HCC. A: FAM83A mRNA levels in normal hepatocytes and HCC cell lines based on the TCGA (http://ualcan.path.uab.edu/ index.html) database. B: Western blot analysis of FAM83A expression levels in normal hepatocytes and HCC cell lines. C: FAM83A expression was measured via immunohistochemical staining in HCC and paracarcinoma tissues. D, E: Kaplan-Meier survival analysis for overall survival and progression-free survival of 82 HCC patients based on the FAM83A expression data. All experiments were performed in triplicate.

Article Snippet: Diluted primary antibody of anti-rabbit FAM83A (1:50; Cat. No. 20618-1-AP, Proteintech, Chicago, USA) was used.

Techniques: Western Blot, Expressing, Immunohistochemical staining, Staining

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Positive feedback loop of FAM83A/PI3K/AKT/c-Jun induces migration, invasion and metastasis in hepatocellular carcinoma.

doi: 10.1016/j.biopha.2019.109780

Figure Lengend Snippet: Fig. 2. Overexpression of FAM83A promoted HCC cell invasion and migration in vitro. A: Representative images of HCC cells stably transfected with fluorescently labeled lentivirus under bright-field and fluorescence microscopy. Scale bar: 250 μm. B-G: Both Bel-7404 and SMMC-7721 cells were transfected with negative control (NC) lentivirus or FAM83A lentivirus. B: RT-qPCR analysis of FAM83A mRNA expression. C: Protein levels of FAM83A, PI3K, AKT, p-PI3K, p-AKT, c-JUN, E- cadherin, N-cadherin and Vimentin were analyzed via western blot. D, E: A wound-healing assay was performed to analyze the migratory abilities of Bel-7404 and SMMC-7721 cells for 0, 24, 48 or 72 h and detected via wound-healing assays. Scale bar: 250 μm. F, G: Migration and invasion abilities of Bel-7404 and SMMC-772 were determined using transwell and Boyden chamber assays. Scale bar: 250 μm. Data are presented as mean ± s.d. from three independent experiments. *P < 0.05 versus control; **P < 0.01; ***P < 0.001.

Article Snippet: Diluted primary antibody of anti-rabbit FAM83A (1:50; Cat. No. 20618-1-AP, Proteintech, Chicago, USA) was used.

Techniques: Over Expression, Migration, In Vitro, Stable Transfection, Transfection, Labeling, Microscopy, Negative Control, Quantitative RT-PCR, Expressing, Western Blot, Wound Healing Assay, Control

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Positive feedback loop of FAM83A/PI3K/AKT/c-Jun induces migration, invasion and metastasis in hepatocellular carcinoma.

doi: 10.1016/j.biopha.2019.109780

Figure Lengend Snippet: Fig. 3. FAM83A silencing by siRNAs inhibited HCC cell invasion and migration in vitro. A-F: Both Bel-7404 and SMMC-7721 cells were transfected with NC siRNA or FAM83A siRNA. A: RT-qPCR analysis of FAM83A mRNA expression. B: Protein levels of FAM83A, PI3K, AKT, p-PI3K, p-AKT, c-JUN, E-cadherin, N-cadherin and Vimentin were analyzed via western blot. C, D: A wound-healing assay was performed to analyze the migration abilities of Bel-7404 and SMMC-7721 cells for 0, 24, 48 or 72 h and detected via wound-healing assays. Scale bar: 250 μm. E, F: Migration and invasion abilities of Bel-7404 and SMMC-7721 were determined via transwell and Boyden chamber assays. Scale bar: 250 μm. Data are presented as mean ± s.d. from three independent experiments. *P < 0.05 versus control; **P < 0.01; ***P < 0.001.

Article Snippet: Diluted primary antibody of anti-rabbit FAM83A (1:50; Cat. No. 20618-1-AP, Proteintech, Chicago, USA) was used.

Techniques: Migration, In Vitro, Transfection, Quantitative RT-PCR, Expressing, Western Blot, Wound Healing Assay, Control

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Positive feedback loop of FAM83A/PI3K/AKT/c-Jun induces migration, invasion and metastasis in hepatocellular carcinoma.

doi: 10.1016/j.biopha.2019.109780

Figure Lengend Snippet: Fig. 4. FAM83A induced sorafenib resistance. A, C: Bel-7404 and SMMC-7721 cells were transfected with NC lentivirus or FAM83A lentivirus. Sorafenib IC50 values of the HCC cell lines. B, D: Bel-7404 and SMMC-7721 cells were transfected with NC siRNA or FAM83A siRNA. Sorafenib IC50 values of the HCC cell lines. Data are presented as mean ± s.d. from three independent experiments. *P < 0.05 versus control; **P < 0.01; ***P < 0.001.

Article Snippet: Diluted primary antibody of anti-rabbit FAM83A (1:50; Cat. No. 20618-1-AP, Proteintech, Chicago, USA) was used.

Techniques: Transfection, Control

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Positive feedback loop of FAM83A/PI3K/AKT/c-Jun induces migration, invasion and metastasis in hepatocellular carcinoma.

doi: 10.1016/j.biopha.2019.109780

Figure Lengend Snippet: Fig. 5. FAM83A promoted metastatic capacity of HCC cells in vivo. A: The pulmonary metastasis model was adopted to evaluate the effect of FAM83A on HCC cell metastasis. B: The number of mice with pulmonary metastasis was calculated in each group. C: Statistics of pulmonary metastatic nodules. *P < 0.05 versus control. D: Representative pictures of lung metastatic nodules under bright-field microscopy (BF), fluorescence microscopy showing green fluorescent protein expression (GFP) and H&E staining. Scale bar: 2 mm.

Article Snippet: Diluted primary antibody of anti-rabbit FAM83A (1:50; Cat. No. 20618-1-AP, Proteintech, Chicago, USA) was used.

Techniques: In Vivo, Control, Microscopy, Expressing, Staining

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Positive feedback loop of FAM83A/PI3K/AKT/c-Jun induces migration, invasion and metastasis in hepatocellular carcinoma.

doi: 10.1016/j.biopha.2019.109780

Figure Lengend Snippet: Fig. 6. c-JUN promoted FAM83A expression by binding to its promoter region. A: Schematic diagram of the promoter regions of FAM83A with the putative c-JUN transcription factor-binding sites (P1–P4). b–c Bel-7404 and SMMC-7721 cells were transfected with NC or c-JUN plasmids. B: The mRNA levels of c-JUN and FAM83A were measured via RT-qPCR. C: The protein levels of c-JUN and FAM83A were measured via western blot. D: Amplification of the c-JUN-binding sites, P1 and P3, after ChIP using c-JUN antibody. E: EMSA and supershift assays of c-JUN binding to the FAM83A promoter in Bel-7404 and SMMC-7721 cells. The free probe of labeled c-JUN was run in lane 1 as a control. A 100-fold excess of unlabeled c-JUN-WT was used to compete with c-JUN binding (lane 2 compared with lane 6). A 100-fold excess of unlabeled mutated c-JUN-P1, c-JUN-P3 or c-JUN-P1+P3 was used to compete with binding of the respective labeled probes (lanes 3–5 compared with lane 6). A supershift assay (lane 7) was performed using an anti-c-JUN antibody. F: FAM83A activated the PI3K/AKT signal pathway and induced EMT signaling, further promoting migration, invasion, metastasis and sorafenib resistance in HCC cells. c-JUN, a downstream protein of the PI3K/AKT pathway, promoted FAM83A expression by binding to the FAM83A promoter region. Data are presented as mean ± s.d. from three independent experiments. *P < 0.05 versus control; **P < 0.01; ***P < 0.001.

Article Snippet: Diluted primary antibody of anti-rabbit FAM83A (1:50; Cat. No. 20618-1-AP, Proteintech, Chicago, USA) was used.

Techniques: Expressing, Binding Assay, Transfection, Quantitative RT-PCR, Western Blot, Labeling, Control, Migration

Journal: Frontiers in Oncology

Article Title: FAM83A Promotes Lung Cancer Progression by Regulating the Wnt and Hippo Signaling Pathways and Indicates Poor Prognosis

doi: 10.3389/fonc.2020.00180

Figure Lengend Snippet: Expression of FAM83A in lung cancers and its correlation with prognosis. Expression levels of FAM83A in lung cancers and normal lung tissues, and their significant relation to the prognosis of patients with lung cancer. Box plots of FAM83A expression levels in lung adenocarcinomas (LUADs) (A) and lung squamous cell carcinomas (LUSC) (B) compared to normal lung tissues, which were retrieved from the UALCAN database. Kaplan–Meier curves of FAM83A expression in LUAD (C) and LUSC (D) , as retrieved from the UALCAN database.

Article Snippet: The membrane was blocked with 5% non-fat milk for 2 h and incubated overnight at 4°C with antibodies against FAM83A (1:700, Origene, #TA335330), cyclin D1 (1:100, #sc-8396; Santa Cruz Biotechnology, Santa Cruz, CA, USA), MMP7 (1:100, #sc-515703; Santa Cruz Biotechnology), GAPDH (1:2,000, #sc-47724; Santa Cruz Biotechnology), β-catenin (1:500, #8480; Cell Signaling Technology, Danvers, MA, USA), active β-catenin (1:500, #8814; Cell Signaling Technology), E-cadherin (1:500, #3195; Cell Signaling Technology), GSK3β (1:500, #9832; Cell Signaling Technology), phosphorylated yes-associated protein (p-YAP) (1:500, #13008; Cell Signaling Technology), cyclin E (1:500, #11554-1-AP; Proteintech, Chicago, IL, USA), Twist (1:500, #25465-1-AP; Proteintech), Snail (1:500, #13099-1-AP; Proteintech), vimentin (1:500, #10366-1-AP; Proteintech), YAP (1:500, #13584-1-AP; Proteintech), phosphorylated GSK3β (p-GSK3β) (1:500, #14850-1-AP; Proteintech), LATS1 (1:500, #17049-1-AP; Proteintech), MST1 (1:500, #22245-1-AP; Proteintech), CTGF (1:500, #23936-1-AP; Proteintech), and c-Myc (1:500, #551101; BD Biosciences, San Jose, CA, USA).

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: FAM83A Promotes Lung Cancer Progression by Regulating the Wnt and Hippo Signaling Pathways and Indicates Poor Prognosis

doi: 10.3389/fonc.2020.00180

Figure Lengend Snippet: Effects of FAM83A on the proliferation, colony formation, and invasion of lung cancer cells. (A,B) The expression level of FAM83A of H1299 and A549 cells transfected with FAM83A or SiFAM83A. (C) The cell growth curve of H1299 and A549 cells transfected with FAM83A or SiFAM83A, and their control cells. (D–G) Representative images of the colony formation assay for H1299 and A549 cells transfected with FAM83A (D) or SiFAM83A (F) , and their control cells. The number of colonies formed by each group is shown in the histogram (E,G) . Representative images of the Matrigel invasion assay for H1299 and A549 cells transfected with FAM83A (H) or SiFAM83A (J) and their control cells. The invasive cell number for each group is shown in the histogram (I,K) . * P < 0.05, ** P < 0.01, *** P < 0.001, NC, negative control cells; FAM83A, cells transfected with FAM83A; SiNC, cells interfered with control SiRNA; SiFAM83A, cells interfered with SiFAM83A.

Article Snippet: The membrane was blocked with 5% non-fat milk for 2 h and incubated overnight at 4°C with antibodies against FAM83A (1:700, Origene, #TA335330), cyclin D1 (1:100, #sc-8396; Santa Cruz Biotechnology, Santa Cruz, CA, USA), MMP7 (1:100, #sc-515703; Santa Cruz Biotechnology), GAPDH (1:2,000, #sc-47724; Santa Cruz Biotechnology), β-catenin (1:500, #8480; Cell Signaling Technology, Danvers, MA, USA), active β-catenin (1:500, #8814; Cell Signaling Technology), E-cadherin (1:500, #3195; Cell Signaling Technology), GSK3β (1:500, #9832; Cell Signaling Technology), phosphorylated yes-associated protein (p-YAP) (1:500, #13008; Cell Signaling Technology), cyclin E (1:500, #11554-1-AP; Proteintech, Chicago, IL, USA), Twist (1:500, #25465-1-AP; Proteintech), Snail (1:500, #13099-1-AP; Proteintech), vimentin (1:500, #10366-1-AP; Proteintech), YAP (1:500, #13584-1-AP; Proteintech), phosphorylated GSK3β (p-GSK3β) (1:500, #14850-1-AP; Proteintech), LATS1 (1:500, #17049-1-AP; Proteintech), MST1 (1:500, #22245-1-AP; Proteintech), CTGF (1:500, #23936-1-AP; Proteintech), and c-Myc (1:500, #551101; BD Biosciences, San Jose, CA, USA).

Techniques: Expressing, Transfection, Control, Colony Assay, Invasion Assay, Negative Control

Journal: Frontiers in Oncology

Article Title: FAM83A Promotes Lung Cancer Progression by Regulating the Wnt and Hippo Signaling Pathways and Indicates Poor Prognosis

doi: 10.3389/fonc.2020.00180

Figure Lengend Snippet: Expression of Wnt- and EMT-related genes under the regulation of FAM83A in lung cancer cells. Expression of Wnt signaling pathway- and EMT-related proteins after FAM83A overexpression (A) or knockdown (D) and relative protein levels in H1299 cells (B,E) and A549 cells (C,F) . GAPDH served as an internal control (* P < 0.05, ** P < 0.01, NC, negative control cells; FAM83A, cells transfected with FAM83A; SiNC, cells interfered with control SiRNA; SiFAM83A, cells interfered with SiFAM83A).

Article Snippet: The membrane was blocked with 5% non-fat milk for 2 h and incubated overnight at 4°C with antibodies against FAM83A (1:700, Origene, #TA335330), cyclin D1 (1:100, #sc-8396; Santa Cruz Biotechnology, Santa Cruz, CA, USA), MMP7 (1:100, #sc-515703; Santa Cruz Biotechnology), GAPDH (1:2,000, #sc-47724; Santa Cruz Biotechnology), β-catenin (1:500, #8480; Cell Signaling Technology, Danvers, MA, USA), active β-catenin (1:500, #8814; Cell Signaling Technology), E-cadherin (1:500, #3195; Cell Signaling Technology), GSK3β (1:500, #9832; Cell Signaling Technology), phosphorylated yes-associated protein (p-YAP) (1:500, #13008; Cell Signaling Technology), cyclin E (1:500, #11554-1-AP; Proteintech, Chicago, IL, USA), Twist (1:500, #25465-1-AP; Proteintech), Snail (1:500, #13099-1-AP; Proteintech), vimentin (1:500, #10366-1-AP; Proteintech), YAP (1:500, #13584-1-AP; Proteintech), phosphorylated GSK3β (p-GSK3β) (1:500, #14850-1-AP; Proteintech), LATS1 (1:500, #17049-1-AP; Proteintech), MST1 (1:500, #22245-1-AP; Proteintech), CTGF (1:500, #23936-1-AP; Proteintech), and c-Myc (1:500, #551101; BD Biosciences, San Jose, CA, USA).

Techniques: Expressing, Over Expression, Knockdown, Control, Negative Control, Transfection

Journal: Frontiers in Oncology

Article Title: FAM83A Promotes Lung Cancer Progression by Regulating the Wnt and Hippo Signaling Pathways and Indicates Poor Prognosis

doi: 10.3389/fonc.2020.00180

Figure Lengend Snippet: FAM83A inhibits the activation of the Hippo signaling pathway. Representative results of Western blot analysis (A,D) and relative protein levels in H1299 cells (B,E) and A549 cells (C,F) . GAPDH served as an internal control (* P < 0.05, ** P < 0.01, NC, negative control cells; FAM83A, cells transfected with FAM83A; SiNC, cells interfered with control SiRNA; SiFAM83A, cells interfered with SiFAM83A).

Article Snippet: The membrane was blocked with 5% non-fat milk for 2 h and incubated overnight at 4°C with antibodies against FAM83A (1:700, Origene, #TA335330), cyclin D1 (1:100, #sc-8396; Santa Cruz Biotechnology, Santa Cruz, CA, USA), MMP7 (1:100, #sc-515703; Santa Cruz Biotechnology), GAPDH (1:2,000, #sc-47724; Santa Cruz Biotechnology), β-catenin (1:500, #8480; Cell Signaling Technology, Danvers, MA, USA), active β-catenin (1:500, #8814; Cell Signaling Technology), E-cadherin (1:500, #3195; Cell Signaling Technology), GSK3β (1:500, #9832; Cell Signaling Technology), phosphorylated yes-associated protein (p-YAP) (1:500, #13008; Cell Signaling Technology), cyclin E (1:500, #11554-1-AP; Proteintech, Chicago, IL, USA), Twist (1:500, #25465-1-AP; Proteintech), Snail (1:500, #13099-1-AP; Proteintech), vimentin (1:500, #10366-1-AP; Proteintech), YAP (1:500, #13584-1-AP; Proteintech), phosphorylated GSK3β (p-GSK3β) (1:500, #14850-1-AP; Proteintech), LATS1 (1:500, #17049-1-AP; Proteintech), MST1 (1:500, #22245-1-AP; Proteintech), CTGF (1:500, #23936-1-AP; Proteintech), and c-Myc (1:500, #551101; BD Biosciences, San Jose, CA, USA).

Techniques: Activation Assay, Western Blot, Control, Negative Control, Transfection

Journal: Frontiers in Oncology

Article Title: FAM83A Promotes Lung Cancer Progression by Regulating the Wnt and Hippo Signaling Pathways and Indicates Poor Prognosis

doi: 10.3389/fonc.2020.00180

Figure Lengend Snippet: FAM83A enhances the proliferative and invasive abilities of lung cancer cells by the Wnt signaling pathway. (A) The cell growth curve of H1299 cells transfected with FAM83A, and their control cells were added with DMSO or XAV-939. (B,C) Representative images of the colony formation assay for H1299 cells transfected with FAM83A, and their control cells were added with DMSO or XAV-939 (B) . The number of colonies formed by each group is shown in the histogram (C) . (D,E) Representative images of the Matrigel invasion assay for H1299 cells transfected with FAM83A, and their control cells were added with DMSO or XAV-939 (D) . The invasive cell number for each group is shown in the histogram (E) . * P < 0.05, ** P < 0.01, *** P < 0.001, NC, negative control cells; FAM83A, cells transfected with FAM83A; DMSO, dimethyl sulfoxide; XAV-939, an inhibitor of Wnt/β-catenin signaling; *NC+DMSO vs. FAM83A+DMSO; *FAM83A+DMSO vs. FAM83A+XAV-939.

Article Snippet: The membrane was blocked with 5% non-fat milk for 2 h and incubated overnight at 4°C with antibodies against FAM83A (1:700, Origene, #TA335330), cyclin D1 (1:100, #sc-8396; Santa Cruz Biotechnology, Santa Cruz, CA, USA), MMP7 (1:100, #sc-515703; Santa Cruz Biotechnology), GAPDH (1:2,000, #sc-47724; Santa Cruz Biotechnology), β-catenin (1:500, #8480; Cell Signaling Technology, Danvers, MA, USA), active β-catenin (1:500, #8814; Cell Signaling Technology), E-cadherin (1:500, #3195; Cell Signaling Technology), GSK3β (1:500, #9832; Cell Signaling Technology), phosphorylated yes-associated protein (p-YAP) (1:500, #13008; Cell Signaling Technology), cyclin E (1:500, #11554-1-AP; Proteintech, Chicago, IL, USA), Twist (1:500, #25465-1-AP; Proteintech), Snail (1:500, #13099-1-AP; Proteintech), vimentin (1:500, #10366-1-AP; Proteintech), YAP (1:500, #13584-1-AP; Proteintech), phosphorylated GSK3β (p-GSK3β) (1:500, #14850-1-AP; Proteintech), LATS1 (1:500, #17049-1-AP; Proteintech), MST1 (1:500, #22245-1-AP; Proteintech), CTGF (1:500, #23936-1-AP; Proteintech), and c-Myc (1:500, #551101; BD Biosciences, San Jose, CA, USA).

Techniques: Transfection, Control, Colony Assay, Invasion Assay, Negative Control

Journal: Frontiers in Oncology

Article Title: FAM83A Promotes Lung Cancer Progression by Regulating the Wnt and Hippo Signaling Pathways and Indicates Poor Prognosis

doi: 10.3389/fonc.2020.00180

Figure Lengend Snippet: GSK3β and the Wnt signaling pathway contribute to FAM83A induced YAP activation. Expression of YAP and downstream proteins after FAM83A overexpression and adding Wnt inhibitor (A) or knocking down and adding GSK-3α/β inhibitor (B) in H1299 cells. Relative expression levels of proteins for each group are shown in the histogram (C,D) . * P < 0.05, ** P < 0.01, NC, negative control cells; FAM83A, cells transfected with FAM83A; SiNC, cells interfered with control SiRNA; SiFAM83A, cells interfered with SiFAM83A; XAV-939, an inhibitor of Wnt/β-catenin signaling; CHIR-99021, a GSK-3α/β inhibitor.

Article Snippet: The membrane was blocked with 5% non-fat milk for 2 h and incubated overnight at 4°C with antibodies against FAM83A (1:700, Origene, #TA335330), cyclin D1 (1:100, #sc-8396; Santa Cruz Biotechnology, Santa Cruz, CA, USA), MMP7 (1:100, #sc-515703; Santa Cruz Biotechnology), GAPDH (1:2,000, #sc-47724; Santa Cruz Biotechnology), β-catenin (1:500, #8480; Cell Signaling Technology, Danvers, MA, USA), active β-catenin (1:500, #8814; Cell Signaling Technology), E-cadherin (1:500, #3195; Cell Signaling Technology), GSK3β (1:500, #9832; Cell Signaling Technology), phosphorylated yes-associated protein (p-YAP) (1:500, #13008; Cell Signaling Technology), cyclin E (1:500, #11554-1-AP; Proteintech, Chicago, IL, USA), Twist (1:500, #25465-1-AP; Proteintech), Snail (1:500, #13099-1-AP; Proteintech), vimentin (1:500, #10366-1-AP; Proteintech), YAP (1:500, #13584-1-AP; Proteintech), phosphorylated GSK3β (p-GSK3β) (1:500, #14850-1-AP; Proteintech), LATS1 (1:500, #17049-1-AP; Proteintech), MST1 (1:500, #22245-1-AP; Proteintech), CTGF (1:500, #23936-1-AP; Proteintech), and c-Myc (1:500, #551101; BD Biosciences, San Jose, CA, USA).

Techniques: Activation Assay, Expressing, Over Expression, Negative Control, Transfection, Control

Journal: Frontiers in Oncology

Article Title: FAM83A Promotes Lung Cancer Progression by Regulating the Wnt and Hippo Signaling Pathways and Indicates Poor Prognosis

doi: 10.3389/fonc.2020.00180

Figure Lengend Snippet: A proposed model to illustrate the role of FAM83A in the Wnt and Hippo signaling pathways. GSK3β phosphorylates β-catenin and results in the degradation of β-catenin, which inhibits the activity of the Wnt signaling pathway. FAM83A can inhibit GSK3β activity and increase the level of active unphosphorylated β-catenin; active β-catenin then transports into the nucleus and activates the Wnt signaling pathway. Meanwhile, similar to β-catenin, FAM83A may also inhibit the phosphorylation and degradation of YAP, which is induced by the Hippo signaling pathway and enhance the activity of YAP through repressing GSK3β. In addition, FAM83A could downregulate the upstream MST to inhibit the activation of the Hippo signaling pathway. Figures were produced using Servier Medical Art ( https://smart.servier.com ).

Article Snippet: The membrane was blocked with 5% non-fat milk for 2 h and incubated overnight at 4°C with antibodies against FAM83A (1:700, Origene, #TA335330), cyclin D1 (1:100, #sc-8396; Santa Cruz Biotechnology, Santa Cruz, CA, USA), MMP7 (1:100, #sc-515703; Santa Cruz Biotechnology), GAPDH (1:2,000, #sc-47724; Santa Cruz Biotechnology), β-catenin (1:500, #8480; Cell Signaling Technology, Danvers, MA, USA), active β-catenin (1:500, #8814; Cell Signaling Technology), E-cadherin (1:500, #3195; Cell Signaling Technology), GSK3β (1:500, #9832; Cell Signaling Technology), phosphorylated yes-associated protein (p-YAP) (1:500, #13008; Cell Signaling Technology), cyclin E (1:500, #11554-1-AP; Proteintech, Chicago, IL, USA), Twist (1:500, #25465-1-AP; Proteintech), Snail (1:500, #13099-1-AP; Proteintech), vimentin (1:500, #10366-1-AP; Proteintech), YAP (1:500, #13584-1-AP; Proteintech), phosphorylated GSK3β (p-GSK3β) (1:500, #14850-1-AP; Proteintech), LATS1 (1:500, #17049-1-AP; Proteintech), MST1 (1:500, #22245-1-AP; Proteintech), CTGF (1:500, #23936-1-AP; Proteintech), and c-Myc (1:500, #551101; BD Biosciences, San Jose, CA, USA).

Techniques: Protein-Protein interactions, Activity Assay, Phospho-proteomics, Activation Assay, Produced

Journal: Oncogenesis

Article Title: FAM83A is amplified and promotes cancer stem cell-like traits and chemoresistance in pancreatic cancer

doi: 10.1038/oncsis.2017.3

Figure Lengend Snippet: FAM83A is overexpressed in pancreatic cancer cell lines and primary human pancreatic cancer. ( a ) FAM83A mRNA expression in a published microarray data set (NCBI/GEO/GSE16515; contains 16 normal and 36 pancreatic tumor samples). ( b ) Kaplan–Meier analysis of overall (left) or disease-free (right) survival for patients with pancreatic cancer in the TCGA data set with low vs high FAM83A expression; * P <0.05. ( c , d ) Western blotting analysis of FAM83A expression in 2 primary normal human pancreatic duct epithelial cell (HPDEC) lines and 10 cultured pancreatic cancer cell lines ( c ) and 10 primary pancreatic cancer tissues (T) and the matched adjacent non-tumor tissues (ANT) ( d ); α-tubulin was used as protein loading control. ( e ) Immunohistochemical staining showing FAM83A protein expression was upregulated in human pancreatic cancer specimens compared with normal pancreatic tissues. ( f ) Kaplan–Meier overall (left) and disease-free (right) survival curves for patients with pancreatic cancer with low vs high FAM83A expression ( n =103; P <0.001, log-rank test).

Article Snippet: Rabbit anti- FAM83A (1:500; Sigma) was incubated with the sections overnight at 4 °C.

Techniques: Expressing, Microarray, Western Blot, Cell Culture, Immunohistochemical staining, Staining

Journal: Oncogenesis

Article Title: FAM83A is amplified and promotes cancer stem cell-like traits and chemoresistance in pancreatic cancer

doi: 10.1038/oncsis.2017.3

Figure Lengend Snippet: Upregulation of FAM83A promotes pancreatic CSC-like traits in vitro. ( a ) Gene set enrichment analysis (GSEA) plot showing positive correlations between high FAM83A expression and stem cell gene signatures (LIM_MAMMARY_STEM_CELL_UP; GAL_LEUKEMIC_STEM_CELL_UP; PECE_ MAMMARY_STEM_CELL_UP) in a published pancreatic cancer data set (GSE16515). ( b ) Western blotting analysis of FAM83A expression in PANC-1 and CFPAC-1 pancreatic adenocarcinoma cells stably expressing FAM83A cDNA; α-tubulin was used as loading control. ( c ) Flow cytometry analysis of the CD133 + population in the indicated cells. ( d ) Representative images of tumorspheres formed by the indicated cells (left); scale bar: 100 μm. Histograms (right) showing the mean number of spheres formed. ( e ) Hoechst 33342 dye exclusion assay showing FAM83A overexpression increased the number of SP + cells. ( f ) Effects of chemotherapeutic drugs on the indicated pancreatic cancer cells. Pancreatic cancer cells were seeded in culture plates and incubated with and without 50 μ M gemcitabine and 5-FU for 24 or 72 h. Apoptotic cells were measured by FACS analysis. Each bar is the mean±s.d. of three independent experiments; * P <0.05.

Article Snippet: Rabbit anti- FAM83A (1:500; Sigma) was incubated with the sections overnight at 4 °C.

Techniques: In Vitro, Expressing, Western Blot, Stable Transfection, Flow Cytometry, Exclusion Assay, Over Expression, Incubation

Journal: Oncogenesis

Article Title: FAM83A is amplified and promotes cancer stem cell-like traits and chemoresistance in pancreatic cancer

doi: 10.1038/oncsis.2017.3

Figure Lengend Snippet: Silencing FAM83A inhibits pancreatic CSC-like traits in vitro . ( a ) Western blotting analysis of FAM83A expression in PANC-1 and CFPAC-1 pancreatic cancer cells stably expressing FAM83A shRNAs. ( b ) Flow cytometry analysis of the CD133 + population in the indicated cells. ( c ) Representative images of the tumorspheres formed by the indicated cells (left). Scale bar: 100 μm. Histograms (right) showing the mean number of spheres formed. ( d ) Hoechst 33342 dye exclusion assay showing that silencing FAM83A decreased the number of SP + cells. ( e ) Effects of chemotherapeutic drugs on pancreatic cancer cells. Pancreatic cancer cells were seeded in culture plates and incubated with and without 50 μ M gemcitabine and 5-FU for 24 and 72 h. Apoptotic cells were measured by FACS analysis. Each bar represents the mean±s.d. of three independent experiments; * P <0.05.

Article Snippet: Rabbit anti- FAM83A (1:500; Sigma) was incubated with the sections overnight at 4 °C.

Techniques: In Vitro, Western Blot, Expressing, Stable Transfection, Flow Cytometry, Exclusion Assay, Incubation

Journal: Oncogenesis

Article Title: FAM83A is amplified and promotes cancer stem cell-like traits and chemoresistance in pancreatic cancer

doi: 10.1038/oncsis.2017.3

Figure Lengend Snippet: FAM83A promotes pancreatic cancer tumorigenesis and chemoresistance in vivo. ( a ) Tumor formation rate for different dilutions and estimated percentages of CSCs. ( b ) Representative images of tumor-bearing mice in each group. ( c ) Representative tumor growth curves of xenografts derived from each group treated with gemcitabine (80 mg/kg). ( d ) Tumor size; results are mean±s.d. of three independent experiments; * P <0.05.

Article Snippet: Rabbit anti- FAM83A (1:500; Sigma) was incubated with the sections overnight at 4 °C.

Techniques: In Vivo, Derivative Assay

Journal: Oncogenesis

Article Title: FAM83A is amplified and promotes cancer stem cell-like traits and chemoresistance in pancreatic cancer

doi: 10.1038/oncsis.2017.3

Figure Lengend Snippet: Overexpression of FAM83A activates multiple CSC-associated signaling pathways. ( a ) Gene set enrichment analysis (GSEA) plot showing the positive correlations between high FAM83A expression and TGF-β/Smad and Wnt/β-catenin pathway gene signatures (KARAKAS_TGFB1_SIGNALING; KARLSSON_TGFB_TARGETS_UP; TRACTOME_SIGNALING_BY WNT; LABBE_TARGETSOF TGFB1_AND_WNT3A_UP) in published TCGA data sets ( n =178). ( b ) Relative luciferase activity of TGF-β reporter and TOP/FOP reporter genes. ( c ) Western blotting (WB) analysis of p-Smad3 (Ser423/425) and nuclear β-catenin in the indicated cells; α-tubulin was used as the loading control. ( d ) WB of p-Smad3 and nuclear β-catenin expression in ten freshly isolated pancreatic cancer tissues; α-tubulin was used as the loading control. ( e ) Histograms (right) of the mean number of spheres formed by FAM83A-transduced cells treated with a TGF-β inhibitor and/or β-catenin inhibitor. Each bar represents the mean±s.d. of three independent experiments; * P <0.05.

Article Snippet: Rabbit anti- FAM83A (1:500; Sigma) was incubated with the sections overnight at 4 °C.

Techniques: Over Expression, Expressing, Luciferase, Activity Assay, Western Blot, Isolation

Journal: Oncogenesis

Article Title: FAM83A is amplified and promotes cancer stem cell-like traits and chemoresistance in pancreatic cancer

doi: 10.1038/oncsis.2017.3

Figure Lengend Snippet: Aberrant FAM83A contributes to progression of various types of cancer. ( a ) Analysis of FAM83A CNV in TCGA data sets revealed the FAM83A locus is amplified in 35.5% of pancreatic cancer samples. ( b ) FAM83A gene CNV and corresponding mRNA expression in a TCGA pancreatic cancer data set ( P <0.05, n =177). ( c ) FAM83A gene CNV and corresponding mRNA expression in a TCGA pancreatic cancer data set ( P <0.05, n =177). ( d ) Kaplan–Meier analysis of overall or disease-free survival for patients with low or high FAM83A expression.

Article Snippet: Rabbit anti- FAM83A (1:500; Sigma) was incubated with the sections overnight at 4 °C.

Techniques: Amplification, Expressing