Journal: Oxidative medicine and cellular longevity

Article Title: The Interaction of Apelin and FGFR1 Ameliorated the Kidney Fibrosis through Suppression of TGF β -Induced Endothelial-to-Mesenchymal Transition.

doi: 10.1155/2023/5012474

Figure Lengend Snippet: Figure 1: Proximity between apelin and FGFR1 suppresses the TGFβ/Smad signaling pathway in HMVECs. (a) HMVECs were treated with TGFβ2 (5 ng/mL) or N-FGFR1 (1.5 μg/mL) for 48 h with or without preincubation with apelin (100 nM) for 2 h. The proximity between apelin and FGFR1 was then analyzed by the Duolink In Situ Assay. For each slide, images at a ×400 original magnification were obtained from six different areas. The scale bar is 60 μm in each panel. (b) HMVECs were treated with TGFβ2 (5 ng/mL) or with apelin (100 nM) for 48 h, and the p-Smad3, TGFβR1, TGFβR2, and FGFR1 levels were analyzed by western blot. Densitometric analysis of the p-Smad3/Smad3, TGFβR1/β-actin, TGFβR2/β-actin, and FGFR1/β-actin levels from each group (n = 5) was analyzed. (c) HMVECs were treated with or without apelin (100 nM) for 48 h, and the FGFR1 levels were analyzed by western blot. Densitometric analysis of the FGFR1/GADPH levels from each group (n = 5) was analyzed.

Article Snippet: Human neutralizing FGFR1 (MAB765) was obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: In Situ, Western Blot

Journal: Oxidative medicine and cellular longevity

Article Title: The Interaction of Apelin and FGFR1 Ameliorated the Kidney Fibrosis through Suppression of TGF β -Induced Endothelial-to-Mesenchymal Transition.

doi: 10.1155/2023/5012474

Figure Lengend Snippet: Figure 4: CEBPA knockdown promotes TGFβ-mediated EndMT. (a) HMVECs were transfected with or without CEBPA siRNA for 48 h in the presence or absence of TGFβ2, and the VE-cadherin, α-SMA, vimentin, and SM22α levels were analyzed by western blot. Densitometric analysis of the VE-cadherin/GADPH, α-SMA/GADPH, vimentin/GADPH, and SM22α/GADPH levels from each group (n = 5) was analyzed. (b) HMVECs were transfected with CEBPA siRNA for 48 h in the presence or absence of TGFβ2 and apelin, and the VE- cadherin, α-SMA, vimentin, and SM22α levels were analyzed by western blot. Densitometric analysis of the VE-cadherin/GADPH, α- SMA/GADPH, vimentin/GADPH, and SM22α/GADPH levels from each group (n = 5) was analyzed. (c) HMVECs were treated with TGFβ2 for 15 min or 48 h with or without preincubation with apelin for 2 h, and the CEBPA levels were analyzed by western blot. Densitometric analysis of the CEBPA/GADPH level from each group (n = 5) was analyzed. (d) HMVECs were treated with N-FGFR1 for 48 h or 15 min in the presence or absence of apelin, and the CEBPA levels was analyzed by western blot. Densitometric analysis of the CEBPA/GADPH level from each group (n = 5) was analyzed. Immunofluorescence analysis of CD31 (e) and α-SMA (f) coexpression in HUVECs following TGFβ2 or/and apelin or/and CEBPA siRNA treatment. For each slide, images of six different fields of view at ×200 magnification were evaluated. The scale bar is 50 μm in each panel.

Article Snippet: Human neutralizing FGFR1 (MAB765) was obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Knockdown, Transfection, Western Blot

Journal: PLOS ONE

Article Title: A secreted proteomic footprint for stem cell pluripotency

doi: 10.1371/journal.pone.0299365

Figure Lengend Snippet: Secreted proteins significantly changed in both RNA and protein abundances in both Rebl.PAT and Man-13 cell lines.

Article Snippet: Gels were transferred using the IBlot2 Gel Transfer device (Thermo #IB21001), using iBlot 2 Transfer stacks (nitrocellulose membrane, Thermo #IB23001).Cells were analysed by immunoblotting using the following antibodies: 1/200 Mouse anti-Chromogranin A (Novus Biologicalis, NBP2-44774); 1/100 Rabbit anti-COCH (Abcam, ab170266); 1/500 mouse anti-FGFR1 (R&D systems, MAB658); 1/1000 rabbit anti-Follistatin (Abcam, ab157471); 1/1000 rabbit anti-Entactin/NID1 (Abcam, ab133686); 1:1000 rabbit anti-Neuronal Pentaxtrin 2 (Abcam, ab191563); 1/1000 rabbit anti-Neurotensin (Abcam, ab172114); 1/300 rabbit anti-Olfactomedin-like 3 (Abcam, ab111712); 1/500 mouse anti-Semaphorin 3A (R&D, MAB1250); 1/500 mouse anti-Secreted Frizzled Related Protein 2 (R&D, MAB6838).

Techniques:

Journal: PLOS ONE

Article Title: A secreted proteomic footprint for stem cell pluripotency

doi: 10.1371/journal.pone.0299365

Figure Lengend Snippet: Equal quantities of protein from media supernatant from cells cultured as described in were run on Western blot and probed with antibodies for an E8-enriched marker protein (CHGA, NID1, SEMA3 or NPTX3) and an E6-enriched marker protein (COCH, FGFR1, Follistatin or OLFML3). Membranes were imaged using LICOR Odyssey system.

Article Snippet: Gels were transferred using the IBlot2 Gel Transfer device (Thermo #IB21001), using iBlot 2 Transfer stacks (nitrocellulose membrane, Thermo #IB23001).Cells were analysed by immunoblotting using the following antibodies: 1/200 Mouse anti-Chromogranin A (Novus Biologicalis, NBP2-44774); 1/100 Rabbit anti-COCH (Abcam, ab170266); 1/500 mouse anti-FGFR1 (R&D systems, MAB658); 1/1000 rabbit anti-Follistatin (Abcam, ab157471); 1/1000 rabbit anti-Entactin/NID1 (Abcam, ab133686); 1:1000 rabbit anti-Neuronal Pentaxtrin 2 (Abcam, ab191563); 1/1000 rabbit anti-Neurotensin (Abcam, ab172114); 1/300 rabbit anti-Olfactomedin-like 3 (Abcam, ab111712); 1/500 mouse anti-Semaphorin 3A (R&D, MAB1250); 1/500 mouse anti-Secreted Frizzled Related Protein 2 (R&D, MAB6838).

Techniques: Cell Culture, Western Blot, Marker

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Frequent FGFR1 hotspot alterations in driver-unknown low-grade glioma and mixed neuronal-glial tumors

doi: 10.1007/s00432-021-03906-x

Figure Lengend Snippet: A Distribution of Non-FGFR1 mutations across 476 tumors. Sample numbers are provided on top for each slice. 108 samples without known mutation were selected for FGFR1 testing. B Representative pyrograms for FGFR1 N546K mutation (five cases with c.1638C > A and four with c.1638C > G). C Representative pyrogram for FGFR1 K656E mutation (four cases with c.1966A > G)

Article Snippet: FGFR1 protein expression was detected by immunohistochemistry using a polyclonal antibody raised against phospho-FGFR1 (Tyr653, Tyr654) of human origin (RRID:AB_1500112, #44-1140G, Thermo-Fisher Waltham, MA, USA).

Techniques: Mutagenesis

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Frequent FGFR1 hotspot alterations in driver-unknown low-grade glioma and mixed neuronal-glial tumors

doi: 10.1007/s00432-021-03906-x

Figure Lengend Snippet: Epidemiological data of samples used for comparative FGFR1 pyrosequencing and immunohistochemistry

Article Snippet: FGFR1 protein expression was detected by immunohistochemistry using a polyclonal antibody raised against phospho-FGFR1 (Tyr653, Tyr654) of human origin (RRID:AB_1500112, #44-1140G, Thermo-Fisher Waltham, MA, USA).

Techniques:

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Frequent FGFR1 hotspot alterations in driver-unknown low-grade glioma and mixed neuronal-glial tumors

doi: 10.1007/s00432-021-03906-x

Figure Lengend Snippet: FGFR1 immunohistochemistry

Article Snippet: FGFR1 protein expression was detected by immunohistochemistry using a polyclonal antibody raised against phospho-FGFR1 (Tyr653, Tyr654) of human origin (RRID:AB_1500112, #44-1140G, Thermo-Fisher Waltham, MA, USA).

Techniques:

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Frequent FGFR1 hotspot alterations in driver-unknown low-grade glioma and mixed neuronal-glial tumors

doi: 10.1007/s00432-021-03906-x

Figure Lengend Snippet: Molecular results and frequencies of mutations across diagnosis from full cohort

Article Snippet: FGFR1 protein expression was detected by immunohistochemistry using a polyclonal antibody raised against phospho-FGFR1 (Tyr653, Tyr654) of human origin (RRID:AB_1500112, #44-1140G, Thermo-Fisher Waltham, MA, USA).

Techniques: Biomarker Discovery