Journal: The Journal of Biological Chemistry

Article Title: Reactivation of Mitogen-activated Protein Kinase (MAPK) Pathway by FGF Receptor 3 (FGFR3)/Ras Mediates Resistance to Vemurafenib in Human B-RAF V600E Mutant Melanoma *

doi: 10.1074/jbc.M112.377218

Figure Lengend Snippet: Enhanced FGFR3 activation in vemurafenib-resistant B-RAF V600E melanoma cells. A, phospho-RTK antibody array analysis. Cell lysates from A375, M14, A375-R1, and M14-R cell lines were incubated on RTK antibody array for 16 h and phosphorylation status was determined as described under “Experimental Procedures.” Each RTK antibody is spotted in duplicate. Supplemental Table S2 describes the list of RTKs and the layout of the antibody array. B, confirmation of phospho-FGFR3 levels by Western blot analysis. Protein levels of total and phosho-FGFR3 were assessed using immunoblotting. C, ELISA analysis of secreted FGF2 in the conditioned media obtained from A375, A375-R1, M14, and M14-R cells. ELISA was performed as described in “Experimental Procedures.”

Article Snippet: Cells were cultured for 48 h in growth medium described above, and the conditioned medium samples (cell free culture supernatant) were analyzed for concentrations of human FGF2 using human FGF2 Quantikine ELISA Kit (R&D Systems).

Techniques: Activation Assay, Ab Array, Incubation, Phospho-proteomics, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Materials & Design

Article Title: Multifunctional nanofiber-based dressings in coordination with adipose-derived stem cells for accelerated burn wound healing

doi: 10.1016/j.matdes.2025.113929

Figure Lengend Snippet: Fig. 4. (A) Detection of ADSCs proliferation by ADM coating at different times. Quantification of (B) EGF and (C) FGF2 in ADSCs paracrine products. *** P < 0.001 representing a significant difference as compared with ADM (3 min) and TLWDA.

Article Snippet: Human EGF enzyme-linked immunosorbent assay (ELISA) kit and human FGF2 ELISA kit were purchased from BOSTER (Wuhan, China).

Techniques:

Journal: Regenerative Biomaterials

Article Title: Specific bFGF targeting of KIM-1 in ischemic kidneys protects against renal ischemia-reperfusion injury in rats

doi: 10.1093/rb/rbac029

Figure Lengend Snippet: Purification, bioactivity and targeting capacity of KIT-bFGF. ( A ) Protein structure prediction of recombinant KIT-bFGF and native bFGF. ( B ) Purified KIT-bFGF and native bFGF were detected on the SDS-PAGE gel. ( C ) Purified KIT-bFGF and native bFGF were detected by western blot. ( D ) The bioactivity of KIT-bFGF and native bFGF was detected by MTT assay through the promotion of HSFs proliferation. ( E ) The expression of KIM-1 in HK-2 cells after I/R injury in vitro by western blot. ( F ) The expression of KIM-1 in ischemic kidneys after I/R injury in vivo by western blot. ( G ) Fluorescent observation of targeting capacity of the KIT-bFGF and bFGF proteins labeled with DyLight-747 dye after HK-2 I/R injury for 12 h. The scale bar = 50 μm. ( H ) Cell survival was detected by MTT after KIT-bFGF and bFGF treatment in HK-2 cells after I/R injury in vitro. * P < 0.05.

Article Snippet: And then, extracted proteins and serum were analyzed using a human bFGF ELISA kit (Boster, Wuhan, China) according to the protocol.

Techniques: Purification, Recombinant, SDS Page, Western Blot, MTT Assay, Expressing, In Vitro, In Vivo, Labeling

Journal: Regenerative Biomaterials

Article Title: Specific bFGF targeting of KIM-1 in ischemic kidneys protects against renal ischemia-reperfusion injury in rats

doi: 10.1093/rb/rbac029

Figure Lengend Snippet: KIT-bFGF could target to the ischemic kidney and retain in the kidney after renal I/R injury. ( A ) Quantitative ELISA assay for bFGF in kidney at 6 and 24 h postinjection. At 6 h postinjection, KIT-bFGF = 24.507 ± 2.002 μg/g, bFGF = 13.523 ± 3.704 μg/g and PBS = 8.208 ± 1.562 μg/g. At 24 h postinjection, KIT-bFGF = 29.696 ± 0.682 μg/g, bFGF = 5.618 ± 0.266 μg/g and PBS = 5.572 ± 0.746 μg/g. Data are presented as mean ± SD. N = 6, ** P < 0.01, *** P < 0.001, **** P < 0.0001. ( B ) Quantitative ELISA assay for bFGF in serum. At 6 h postinjection, KIT-bFGF = 9.407 ± 1.849 μg/ml, bFGF = 8.214 ± 0.974 μg/ml and PBS = 12.399 ± 4.141 μg/ml. At 24 h postinjection, KIT-bFGF = 5.315 ± 0.292 μg/ml, bFGF = 5.554 ± 0.410 μg/ml and PBS = 5.873 ± 0.464 μg/ml. Data are presented as mean ± SD. N = 6. ( C , a–d) Animals' imaging for fluorescence DyLight-747 dye labeled KIT-bFGF and bFGF at 6 and 24 h postinjection in vivo by fluorescence imaging system. Ex/Em = 748/771. (e–h) Fluorescence distribution in kidneys after 6 and 24 h administration. Scale bar = 50 μm. N = 6. ( D, E ) Immunofluorescence staining for bFGF (green) and KIM-1 (red) colocalization (yellow) at 6 and 24 h postadministration. Scale bar = 20 μm. Each slice representative of ×400 original magnification. N = 6.

Article Snippet: And then, extracted proteins and serum were analyzed using a human bFGF ELISA kit (Boster, Wuhan, China) according to the protocol.

Techniques: Enzyme-linked Immunosorbent Assay, Imaging, Fluorescence, Labeling, In Vivo, Immunofluorescence, Staining

Journal: Regenerative Biomaterials

Article Title: Specific bFGF targeting of KIM-1 in ischemic kidneys protects against renal ischemia-reperfusion injury in rats

doi: 10.1093/rb/rbac029

Figure Lengend Snippet: KIT-bFGF improved the recovery of renal function after acute renal I/R injury. ( A ) Summary of Scr data for renal function after administration. * P < 0.05. ** P < 0.01. Data are presented as mean ± SD ( B–E ) Histogram of Scr assay for renal function after I/R injury and at 24, 72 h, 2 and 4-week postadministration. * P < 0.05, ** P < 0.01, **** P < 0.0001.

Article Snippet: And then, extracted proteins and serum were analyzed using a human bFGF ELISA kit (Boster, Wuhan, China) according to the protocol.

Techniques:

Journal: Regenerative Biomaterials

Article Title: Specific bFGF targeting of KIM-1 in ischemic kidneys protects against renal ischemia-reperfusion injury in rats

doi: 10.1093/rb/rbac029

Figure Lengend Snippet: KIT-bFGF protected kidney against acute renal I/R injury. ( A–D ) Histopathological staining (H&E) for kidney pathological injury observation. Each slice representative of ×200 original magnification. Scale bar = 50 μm. ( E–H ) Quantitative analysis of renal tubule injury based on H&E staining. All data are expressed as mean ± SD. **** P < 0.0001. N = 8.

Article Snippet: And then, extracted proteins and serum were analyzed using a human bFGF ELISA kit (Boster, Wuhan, China) according to the protocol.

Techniques: Staining

Journal: Regenerative Biomaterials

Article Title: Specific bFGF targeting of KIM-1 in ischemic kidneys protects against renal ischemia-reperfusion injury in rats

doi: 10.1093/rb/rbac029

Figure Lengend Snippet: KIT-bFGF attenuated the renal fibrosis after acute renal I/R injury. ( A, B ) Blocked renal interstitial volume (blue staining and octothorpe) assay by using Masson’s trichrome staining. ×100 original magnification. Scale bar = 100 μm. ( C, D ) Quantitative analysis of the area of renal interstitial fibrosis in obstructed kidneys. All data are expressed as mean ± SD. **** P < 0.0001. N = 8.

Article Snippet: And then, extracted proteins and serum were analyzed using a human bFGF ELISA kit (Boster, Wuhan, China) according to the protocol.

Techniques: Staining

Journal: Regenerative Biomaterials

Article Title: Specific bFGF targeting of KIM-1 in ischemic kidneys protects against renal ischemia-reperfusion injury in rats

doi: 10.1093/rb/rbac029

Figure Lengend Snippet: KIT-bFGF inhabited the apoptosis through the regulation of proapoptotic proteins. ( A–D ) TUNEL staining for apoptosis cells assessment (green). ×200 original magnification, scale bar = 50 μm. ( E–H ) Quantitative of kidney TUNEL-positive cells. All data are expressed as mean ± SD. *** P < 0.001, **** P < 0.0001. N = 8.

Article Snippet: And then, extracted proteins and serum were analyzed using a human bFGF ELISA kit (Boster, Wuhan, China) according to the protocol.

Techniques: TUNEL Assay, Staining

Journal: Regenerative Biomaterials

Article Title: Specific bFGF targeting of KIM-1 in ischemic kidneys protects against renal ischemia-reperfusion injury in rats

doi: 10.1093/rb/rbac029

Figure Lengend Snippet: KIT-bFGF inhabited the apoptosis through the regulation of proapoptotic proteins. ( A–D ) IHC staining for cleaved-Caspase3 after I/R injury and administration. ×200 original magnification, scale bar = 50 μm. ( E–H ) Quantitative of IHC staining for cleaved caspase-3. All data are expressed as mean ± SD. ** P < 0.01, **** P < 0.0001. N = 8. (E) At 24-h postadministration: NORMAL = 0.40 ± 0.52, KIT-bFGF = 1.14 ± 0.69, bFGF = 1.63 ± 0.74, PBS= 2.29 ± 0.49. (F) At 72-h postadministration: KIT-bFGF = 0.75 ± 0.46, bFGF = 1.88 ± 0.35, PBS= 3.13 ± 1.36. (G) At 2-week postadministration: KIT-bFGF = 0.50 ± 0.53, bFGF = 1.63 ± 0.52, PBS= 5.25 ± 1.49. (H) At 4-week postadministration: KIT-bFGF = 0.50 ± 0.53, bFGF = 3.00 ± 1.87, PBS= 4.00 ± 1.58. ( I–L ) Western blot assayed the expression of cleaved Caspase-3, Bcl-2, ERK1/2, phospho-ERK1/2, Akt and phospho-Akt at 24, 72 h, 2-and 4-week postadministration. β-actin and GAPDH were used as a control.

Article Snippet: And then, extracted proteins and serum were analyzed using a human bFGF ELISA kit (Boster, Wuhan, China) according to the protocol.

Techniques: Immunohistochemistry, Western Blot, Expressing, Control

Journal: Physiological Research

Article Title: miR-491-5p Inhibits Emilin 1 to Promote Fibroblasts Proliferation and Fibrosis in Gluteal Muscle Contracture via TGF-β1/Smad2 Pathway

doi: 10.33549/physiolres.934804

Figure Lengend Snippet: Role of miR-491-5p/Emilin 1 on TGF-β/Smad2 signaling axis and fibrosis. (A) Cells were transfected with miR-491-5p mimics or mimics control, and Emilin 1 overexpression plasmid or vector control. Western blot assay was used to evaluate the effect of miR-491-5p on TGF-β and Smad2, and β-Actin was loaded as control. (B) Grayscale analysis of bands in Figure A was conducted. (C) Cells were transfected with miR-491-5p mimics or mimics control and Emilin 1 overexpression plasmid or vector control. mRNA levels of downstream fibrotic cytokines Collagen I, Collagen III, a-SMA and Elastin were evaluated using qRT-PCR in each group. (D–F) Cells were transfected with miR-491-5p mimics or mimics control and Emilin 1 overexpression plasmids or vector control. Levels of TGF-β1 (D), TGF-β3 (E) and CTGF (F) were evaluated with ELISA assay in each group. All data analysis above used One-Way ANOVA analysis of variance. Data were shown as the mean ± SD, n = 3. *p<0.05 and **p<0.01.

Article Snippet: In brief, the supernatants of lytic CB tissues or fibroblasts were used to monitor the concentration of fibrogenic cytokines using Emilin 1 (HZ-2997, Shanghai, CN), TGF-β1 kits (88-8350-22, Invitrogen, US), TGF-β3 kits (KA4402, Abnova, US) and CTGF kits (ABIN6730903, Abnova, US).

Techniques: Transfection, Control, Over Expression, Plasmid Preparation, Western Blot, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay