Journal: International Journal of Molecular Sciences

Article Title: Multiparametric Classification of Non-Muscle Invasive Papillary Urothelial Neoplasms: Combining Morphological, Phenotypical, and Molecular Features for Improved Risk Stratification

doi: 10.3390/ijms23158133

Figure Lengend Snippet: Histoscores of cases according to the histological grading.

Article Snippet: The electrocharged slides were stained with CK20 (M7019, Dako Cytomation, Glostrup, Denmark), p16 (06594441001, CINtech, Roche, Manheim, Germany) p21 (556431, BD Pharmigen, San Jose, CA, USA), p53 (DO-7, Novocastra, Leica Biosystems, Wetzlar, Germany), MIB-1 (M7240, Dako Cytomation, Glostrup, Denmark), p-HH3 (117C826, IMGENEX, San Diego, CA, USA) and FGFR3 (sc-13121, Santa Cruz Biotechnology, Heidelberg, Germany).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Multiparametric Classification of Non-Muscle Invasive Papillary Urothelial Neoplasms: Combining Morphological, Phenotypical, and Molecular Features for Improved Risk Stratification

doi: 10.3390/ijms23158133

Figure Lengend Snippet: Comparison of protein and mutational status among the clusters.

Article Snippet: The electrocharged slides were stained with CK20 (M7019, Dako Cytomation, Glostrup, Denmark), p16 (06594441001, CINtech, Roche, Manheim, Germany) p21 (556431, BD Pharmigen, San Jose, CA, USA), p53 (DO-7, Novocastra, Leica Biosystems, Wetzlar, Germany), MIB-1 (M7240, Dako Cytomation, Glostrup, Denmark), p-HH3 (117C826, IMGENEX, San Diego, CA, USA) and FGFR3 (sc-13121, Santa Cruz Biotechnology, Heidelberg, Germany).

Techniques: Comparison, Immunohistochemistry

Journal: International Journal of Oncology

Article Title: Expression of fibroblast growth factor receptor 2 IIIc in human uterine cervical intraepithelial neoplasia and cervical cancer

doi: 10.3892/ijo_00000504

Figure Lengend Snippet: Figure 1. Preparation of rabbit polyclonal anti-FGFR2 IIIc antibody. Poly- clonal anti-FGFR2 IIIc antibody reacted with recombinant FGFR 2·(IIIc)/ Fc chimera proteins (upper panel, right lane), while the antibody did not react with recombinant FGFR 2·(IIIb)/Fc chimera proteins (upper panel, left lane). Loading control of anti-human IgG antibody reacted with each recom- binant protein on the reblotted membrane (lower panel).

Article Snippet: The following were purchased: recombinant human FGFR 2·(IIIb)/Fc and FGFR 2·(IIIc)/Fc chimera proteins from R&D Systems, Inc. (Minneapolis, MN, USA); horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG secondary antibodies from American Qualex (San Clemente, CA, USA); rabbit polyclonal anti-human IgG and mouse monoclonal anti-Ki-67 antibodies from Dako Japan (Tokyo, Japan); Immobilon P transfer membrane from Millipore (Yonezawa, Japan); Super Signal West Dura chemiluminescent substrates from Thermo Fisher Scientific, Inc. (Waltham, MA, USA); Alexa 488-conjugated goat antirabbit IgG and pcDNA 3.1(-) or (+) vector from Invitrogen (Carlsbad, CA, USA); RNAiso Plus and a FastPure RNA kit from Takara Bio, Inc. (Shiga, Japan); a digoxigenin (DIG) nucleic acid detection kit and a DIG RNA labeling kit from Roche Diagnostics GmbH (Mannheim, Germany); a High Capacity cDNA Reverse Transcription kit, TaqMan Universal PCR Master Mix and a TaqMan MGB probe for ribosomal eukaryotic 18S rRNA from Applied Biosystems (Foster City, CA, USA); Histofine Simple Stain MAX PO (R) or (M) kits from Nichirei (Tokyo, Japan); Matsunami Adhesive Slide (MAS) and 4-well glass bottom dishes from Matsunami Glass Ind., Ltd. (Osaka, Japan); Vectashield mounting medium containing 4',6-diamidino-2-phenylindole dihydrochloride (DAPI) from Vector Laboratories, Inc. (Burlingame, CA, USA); pIRES2-EGFP vector from Clontech (Palo Alto, CA, USA); geneticin from Gibco BRL (Grand Island, NY, USA); WST-8 cell counting kit from Wako Pure Chemical Industries (Osaka, Japan).

Techniques: Recombinant, Control, Membrane

Journal: European urology oncology

Article Title: Profiling Fibroblast Growth Factor Receptor 3 Expression Based on the Immune Microenvironment in Upper Tract Urothelial Carcinoma.

doi: 10.1016/j.euo.2024.01.013

Figure Lengend Snippet: Fig. 1 – (A) Representative immunohistochemistry staining of FGFR3 classified with Ta, T2, T3, and T4. The field scale bar = 500 lm with a low-power field and the field scale bar = 100 lm with a high-power field. (B) Bar chart describing the distribution of FGFR3 expressions in the pT stages. Dark blue: FGFR3 high rates; light blue: FGFR3 low rates. (C) Kaplan-Meier (K-M) curves comparing the recurrence-free survival rates of UTUC patients in the FGFR3 high and low groups. The log-rank test was used to assess the significance of differences. (D) K-M curves comparing the cancer-specific survival rates of UTUC patients in the FGFR3 high and low groups. The log-rank test was used to assess the significance of differences. FGFR3 = fibroblast growth factor receptor 3; UTUC = upper tract urothelial carcinoma.

Article Snippet: The sections were then incubated at 4 C overnight with an anti-FGFR3 mouse monoclonal antibody (Ab; 1:150 dilution; Origene, Rockville, MD, USA), anti-CD4 Ab (1:500 dilution; Abcam, Please cite this article as: K. Shigeta, K. Matsumoto, S. Kitaoka et al., Profilin Microenvironment in Upper Tract Urothelial Carcinoma, Eur Urol Oncol (202 Cambridge, MA, USA), anti-CD8 Ab (1:100 dilution; igene, Abcam), anti-CD68 mouse monoclonal Ab (1:100 dilution; igene, Abcam), anti-CD163 rabbit monoclonal Ab (1:200 dilution; Abcam), anti-CD204 mouse monoclonal Ab (1:200 dilution; Abcam), and anti–PD-L1 rabbit monoclonal Ab (1:500 dilution; Abcam).

Techniques: Immunohistochemistry, Staining

Journal: Molecular cancer therapeutics

Article Title: Cigarette Smoke Containing Acrolein Contributes to Cisplatin Resistance in Human Bladder Cancers through the Regulation of HER2 Pathway or FGFR3 Pathway.

doi: 10.1158/1535-7163.MCT-21-0725

Figure Lengend Snippet: Figure 3. Analysis of HER2 and FGFR3 sig- naling pathways in acrolein- induced cisplatin-resistant RT4 and T24 clones. Western blot anal- ysis of HER2 and FGFR3 down- stream signaling pathways was performed in RT4 Acr-clone#3 (A) and T24 Acr-clone#1 (B) com- pared with parental cells. mRNA expression of HER2 and FGFR3 was analyzed in RT4 Acr-clone#3 (upper panel of C) and T24 Acr-clone#1 (lower panel of C) compared with parental cells using qRT-PCR assay. Values were pre- sented as the mean SD. Stu- dent’s t tests were used to deter- mine statistical significance, and two-tailed P values are shown. , P < 0.01 compared with paren- tal cells. Dose and time effects of acrolein on HER2 and FGFR3 expression, p38 activation, and ERK activation in RT4 Acr-clone#3 (D) and T24 Acr-clone#1 (E) cells were compared with that of paren- tal cells using Western blot analy- sis. For the dose and time effects, the cells were treated with differ- ent concentrations of acrolein (0–10 mmol/L) for 24 hours or acrolein (7.5 mmol/L) for 3 to 24 hours, respectively. h, hour.

Article Snippet: Briefly, blots were blocked with 5% nonfat milk and hybridized with primary antibodies overnight at 4 C. The antibodies against phosphoHER2/ErbB2 (Tyr1221/1222; 1:1000, Cell Signaling Technology #2249), HER2 (1:1000, Cell Signaling Technology #9212), phosphoFGFR3 (Tyr724; 1:1000, Abcam #ab155960), FGFR3 (1:1000, Novus Biologicals #JM81–10), phospho-p38 (Thr180/Tyr182; 1:1000, Cell Signaling Technology #9211), p38 (1:1000, Cell Signaling Technology #9212), phospho-p44/42MAPK (Erk1/2; Thr202/Tyr204; 1:1000, Cell Signaling Technology #9101), p44/42 MAPK (Erk1/2; 1:1000, Cell Signaling Technology #9102), PARP-1 (1:1000, Cell Signaling Technology #9542), caspase-9 (1:1000, Cell Signaling Technology #9502), caspase-3 (1:1000, Cell Signaling Technology #9662), E-cadherin (1:1000, Cell Signaling Technology #3195), EpCAM (gifted from Prof. Ying Chih Chang from Genomic Research Center at Academia Sinica, Taipei, Taiwan), vimentin (1:1000, Taiclone # TCEA19744), Snail AACRJournals.org Mol Cancer Ther; 21(6) June 2022 1011 D ow nloaded from http://aacrjournals.org/m ct/article-pdf/21/6/1010/3153401/1010.pdf by guest on 28 O ctober 2024 (1:1000, Cell Signaling Technology #C15D3), and GAPDH (1:5000, Cell Signaling Technology #5174).

Techniques: Clone Assay, Western Blot, Protein-Protein interactions, Expressing, Quantitative RT-PCR, Two Tailed Test, Activation Assay

Journal: Molecular cancer therapeutics

Article Title: Cigarette Smoke Containing Acrolein Contributes to Cisplatin Resistance in Human Bladder Cancers through the Regulation of HER2 Pathway or FGFR3 Pathway.

doi: 10.1158/1535-7163.MCT-21-0725

Figure Lengend Snippet: Figure 4. Effect of trastuzumab or PD173074 combined with cisplatin in acrolein-induced cisplatin-resistant RT4 and T24 clones. RT4 Acr-clone#3 was treated with cisplatin (2.5 mg/mL), or cisplatin (2.5 mg/mL) with trastuzumab (1 mg/mL), or trastuzumab (1 mg/mL), or cisplatin (2.5 mg/mL) with PD173074 (5 mmol/L), or PD173074 (5 mmol/L) for 24 hours. T24 Acr-clone#1 was treated with cisplatin (1 mg/mL, 24 hours), or cisplatin (1 mg/mL) with PD173074 (5 mmol/L), or PD173074 (5 mmol/L), or cisplatin (1 mg/mL) with trastuzumab (1 mg/mL), or trastuzumab (1 mg/mL) for 24 hours. Cytotoxicity of trastuzumab combined with cisplatin in RT4 Acr-clone#3 (A) and in T24 Acr-clone#1 (D) and cytotoxicity of PD173074 combined with cisplatin in T24 Acr-clone#1 (G) and RT4 Acr-clone#3 (J) compared with that of the parental cells were analyzed using MTT assays. Values are presented as the mean SD. Student t tests were used to determine statistical significance, and two-tailed P values are shown. , P < 0.05, compared with vehicle treatment, ##, P < 0.01; ###, P < 0.005 compared with cisplatin treatment group. Western blot analysis of apoptotic proteins, including cleavage of PARP, caspase-9, and caspase-3, in RT4 Acr-clone#3 (B, K) and T24 Acr-clone#1 (E, H) cells compared with that of the parental cells was performed. Western blot analysis of HER2 and FGFR3, p38 activation, and ERK activation in RT4 Acr-clone#3 (C, L) and T24 Acr-clone#1 (F, I) cells compared with that of the parental cells was performed. c-Cas.: cleavage form of caspase; pro-Cas.: proform of caspase.

Article Snippet: Briefly, blots were blocked with 5% nonfat milk and hybridized with primary antibodies overnight at 4 C. The antibodies against phosphoHER2/ErbB2 (Tyr1221/1222; 1:1000, Cell Signaling Technology #2249), HER2 (1:1000, Cell Signaling Technology #9212), phosphoFGFR3 (Tyr724; 1:1000, Abcam #ab155960), FGFR3 (1:1000, Novus Biologicals #JM81–10), phospho-p38 (Thr180/Tyr182; 1:1000, Cell Signaling Technology #9211), p38 (1:1000, Cell Signaling Technology #9212), phospho-p44/42MAPK (Erk1/2; Thr202/Tyr204; 1:1000, Cell Signaling Technology #9101), p44/42 MAPK (Erk1/2; 1:1000, Cell Signaling Technology #9102), PARP-1 (1:1000, Cell Signaling Technology #9542), caspase-9 (1:1000, Cell Signaling Technology #9502), caspase-3 (1:1000, Cell Signaling Technology #9662), E-cadherin (1:1000, Cell Signaling Technology #3195), EpCAM (gifted from Prof. Ying Chih Chang from Genomic Research Center at Academia Sinica, Taipei, Taiwan), vimentin (1:1000, Taiclone # TCEA19744), Snail AACRJournals.org Mol Cancer Ther; 21(6) June 2022 1011 D ow nloaded from http://aacrjournals.org/m ct/article-pdf/21/6/1010/3153401/1010.pdf by guest on 28 O ctober 2024 (1:1000, Cell Signaling Technology #C15D3), and GAPDH (1:5000, Cell Signaling Technology #5174).

Techniques: Clone Assay, Two Tailed Test, Western Blot, Activation Assay

Journal: Molecular cancer therapeutics

Article Title: Cigarette Smoke Containing Acrolein Contributes to Cisplatin Resistance in Human Bladder Cancers through the Regulation of HER2 Pathway or FGFR3 Pathway.

doi: 10.1158/1535-7163.MCT-21-0725

Figure Lengend Snippet: Figure 5. Effect of PD173074 combined with cisplatin in acrolein-induced cisplatin-resistant T24 clones using an xenograft mouse model. The animal experimental protocol is described in the Materials and Methods. A, Overall view of tumors formed by T24 Acr-clone#1 cells treated with the vehicle (PBS, 50 mL, i.p. once weekly; and corn oil, 50 mL, i.p. once weekly), cisplatin (5 mg/kg in PBS, 50 mL, i.p. once weekly), cisplatin (5 mg/kg in PBS, 50 mL, i.p. once weekly) combined with PD173074 (10 mg/kg in corn oil, 50 mL, i.p. once weekly), or PD173074 (10 mg/kg in corn oil, 50 mL, i.p. once weekly). Tumor growth curves (B) and body weight (C) of nude mice in different experimental groups (n ¼ 5). D, Representative H&E (top), TUNEL staining (middle), and IHC staining of FGFR3 (bottom) in tumor sections from the vehicle, cisplatin, and/or PD173074 treatment groups. E, Quantification of the TUNEL-positive area shown in the middle panel of (D). The apoptotic tumor cells in the middle panel are stained brown. F, Quantification of the FGFR3-positive area shown in the lower panel of (D). Student t tests were used to determine statistical significance, and two-tailed P values are shown. , P < 0.01; , P < 0.005 compared with vehicle group; #, P < 0.05; ##, P < 0.01 compared with cisplatin alone; &&, P < 0.01; &&&, P < 0.005 compared with cisplatin þ PD173074.

Article Snippet: Briefly, blots were blocked with 5% nonfat milk and hybridized with primary antibodies overnight at 4 C. The antibodies against phosphoHER2/ErbB2 (Tyr1221/1222; 1:1000, Cell Signaling Technology #2249), HER2 (1:1000, Cell Signaling Technology #9212), phosphoFGFR3 (Tyr724; 1:1000, Abcam #ab155960), FGFR3 (1:1000, Novus Biologicals #JM81–10), phospho-p38 (Thr180/Tyr182; 1:1000, Cell Signaling Technology #9211), p38 (1:1000, Cell Signaling Technology #9212), phospho-p44/42MAPK (Erk1/2; Thr202/Tyr204; 1:1000, Cell Signaling Technology #9101), p44/42 MAPK (Erk1/2; 1:1000, Cell Signaling Technology #9102), PARP-1 (1:1000, Cell Signaling Technology #9542), caspase-9 (1:1000, Cell Signaling Technology #9502), caspase-3 (1:1000, Cell Signaling Technology #9662), E-cadherin (1:1000, Cell Signaling Technology #3195), EpCAM (gifted from Prof. Ying Chih Chang from Genomic Research Center at Academia Sinica, Taipei, Taiwan), vimentin (1:1000, Taiclone # TCEA19744), Snail AACRJournals.org Mol Cancer Ther; 21(6) June 2022 1011 D ow nloaded from http://aacrjournals.org/m ct/article-pdf/21/6/1010/3153401/1010.pdf by guest on 28 O ctober 2024 (1:1000, Cell Signaling Technology #C15D3), and GAPDH (1:5000, Cell Signaling Technology #5174).

Techniques: Clone Assay, TUNEL Assay, Staining, Immunohistochemistry, Two Tailed Test

Journal: Journal of Histochemistry and Cytochemistry

Article Title: Enhanced Expression of Fibroblast Growth Factor Receptor 3 IIIc Promotes Human Esophageal Carcinoma Cell Proliferation

doi: 10.1369/0022155415616161

Figure Lengend Snippet: RT-PCR Analysis and Immunohistochemical Analysis of FGFR3IIIc Expression in Esophageal Squamous Cell Carcinoma.

Article Snippet: The cell lysates were electrophoresed through 7.5% polyacrylamide gels and transferred to PVDF membranes (Immobilon-P; Millipore), and were analyzed by immunoblotting with the monoclonal anti-β-actin antibody (Sigma-Aldrich; 1:2000 dilution), anti-FGFR3 antibody (C-15) (Santa Cruz Biotechnology, Dallas, TX; 1:2000 dilution) and monoclonal anti-FGFR3IIIc antibody (cat#MAB7662, lot.GHK411041, R&D Systems; 1:500 dilution).

Techniques: Immunohistochemical staining, Expressing

Journal: Journal of Histochemistry and Cytochemistry

Article Title: Enhanced Expression of Fibroblast Growth Factor Receptor 3 IIIc Promotes Human Esophageal Carcinoma Cell Proliferation

doi: 10.1369/0022155415616161

Figure Lengend Snippet: Enhanced expression of FGFR3IIIc was associated with proliferating esophageal carcinoma (EC) cells. (A) Immunofluorescence staining of FGFR3IIIc (a, d) and SCC-112 (b, e) in non-cancerous mucosa (NCM; N, non-tumor) and esophageal squamous cell carcinoma (ESCC) (T, tumor), which was diagnosed as stage 0. In confocal microscopic images, strong staining of FGFR3IIIc (red) was observed in EC cells (d) but not in normal esophageal epithelium cells (a). FGFR3IIIc-positive cells in ESCC were consistent with SCC-112-positive cells (green) (d, e, f: white arrows). Nuclei were stained with DAPI (blue). (B) Immunofluorescence staining of Ki-67 (red; a, d) and SCC-112 (green; b, e) in NCM (N) and ESCC (T). The strong staining of Ki-67 was observed in EC cells, and Ki67-positive cells were consistent with SCC-112-positive cells in ESCC (d, e, f: white arrows). On the other hand, staining of Ki-67 and SCC-112 were not observed in NCM (a, b, c). (C) Immunofluorescence staining of DAPI (a, d), FGFR3IIIc (b), Ki-67 (e) and merged images (c, f) in ESCC samples. The expression of FGFR3IIIc was detected in the same cells, which also expressed Ki-67 in consecutive sections (c, f; white arrows). Scale, 20 μm.

Article Snippet: The cell lysates were electrophoresed through 7.5% polyacrylamide gels and transferred to PVDF membranes (Immobilon-P; Millipore), and were analyzed by immunoblotting with the monoclonal anti-β-actin antibody (Sigma-Aldrich; 1:2000 dilution), anti-FGFR3 antibody (C-15) (Santa Cruz Biotechnology, Dallas, TX; 1:2000 dilution) and monoclonal anti-FGFR3IIIc antibody (cat#MAB7662, lot.GHK411041, R&D Systems; 1:500 dilution).

Techniques: Expressing, Immunofluorescence, Staining

Journal: Journal of Histochemistry and Cytochemistry

Article Title: Enhanced Expression of Fibroblast Growth Factor Receptor 3 IIIc Promotes Human Esophageal Carcinoma Cell Proliferation

doi: 10.1369/0022155415616161

Figure Lengend Snippet: Expression of FGFR3IIIc was enhanced in esophageal carcinoma cells diagnosed as stage IA (A, B, and C), stage IB (D and E), stage IIIA (F, G and H), but not in normal esophageal epithelium cells. FGFR3IIIc was not stained by anti-FGFR3IIIc antibody after pre-adsorption with the recombinant human FGFR3IIIc Fc chimera (H). Hematoxylin and eosin staining (H&E stain, A, D, and F), FGFR3IIIc immunostaining (B, E, and G), and SCC-112 immunostaining (C). Strong staining for FGFR3IIIc was observed in esophageal carcinoma cells (B, E, and G: white arrows), and FGFR3IIIc expression was clearly restricted in the carcinoma area, with a clear border. Normal esophageal epithelium cells did not express FGFR3IIIc (B, E, and G: black arrows). FGFR3IIIc-positive cells were consistent with SCC-112-positive cells in the nuclei on consecutive sections (C: white arrows). Scale, 20 μm.

Article Snippet: The cell lysates were electrophoresed through 7.5% polyacrylamide gels and transferred to PVDF membranes (Immobilon-P; Millipore), and were analyzed by immunoblotting with the monoclonal anti-β-actin antibody (Sigma-Aldrich; 1:2000 dilution), anti-FGFR3 antibody (C-15) (Santa Cruz Biotechnology, Dallas, TX; 1:2000 dilution) and monoclonal anti-FGFR3IIIc antibody (cat#MAB7662, lot.GHK411041, R&D Systems; 1:500 dilution).

Techniques: Expressing, Staining, Adsorption, Recombinant, Immunostaining

Journal: Journal of Histochemistry and Cytochemistry

Article Title: Enhanced Expression of Fibroblast Growth Factor Receptor 3 IIIc Promotes Human Esophageal Carcinoma Cell Proliferation

doi: 10.1369/0022155415616161

Figure Lengend Snippet: PCR Primer Sequences.

Article Snippet: The cell lysates were electrophoresed through 7.5% polyacrylamide gels and transferred to PVDF membranes (Immobilon-P; Millipore), and were analyzed by immunoblotting with the monoclonal anti-β-actin antibody (Sigma-Aldrich; 1:2000 dilution), anti-FGFR3 antibody (C-15) (Santa Cruz Biotechnology, Dallas, TX; 1:2000 dilution) and monoclonal anti-FGFR3IIIc antibody (cat#MAB7662, lot.GHK411041, R&D Systems; 1:500 dilution).

Techniques: Sequencing

Journal: Journal of Histochemistry and Cytochemistry

Article Title: Enhanced Expression of Fibroblast Growth Factor Receptor 3 IIIc Promotes Human Esophageal Carcinoma Cell Proliferation

doi: 10.1369/0022155415616161

Figure Lengend Snippet: Enhanced expression of FGFR3IIIc accelerated cell proliferation. (A) Cell proliferation was significantly lower (by 15%) in EC-GI-10 cells treated with FGFR3 siRNA (siFGFR3) than in EC-GI-10 cells treated with sicontrol (sicontrol) after 5 days in culture. Cell proliferation in FGFR3IIIc-overexpressed siFGFR3-EC-GI-10 cells (FGFR3IIIc) was significantly higher than that in siFGFR3-EC-GI-10 cells (by 1.3-fold), whereas the overexpression of FGFR3IIIb (FGFR3IIIb) did not significantly affect cell proliferation rates. Cell proliferation was significantly lower (~25%) in EGFP-infected siFGFR3-EC-GI-10 cells (EGFP) than in siFGFR3-EC-GI-10 cells. The parental EC-GI-10 cells were not treated with siRNA (-) and not infected (-). Cell numbers are the mean ± S.E.M. The experiment was performed with n=6 samples, and repeated thrice. (#, p<0.05 versus the sicontrol cells and *, p<0.05 versus the siFGFR3 cells; t-test, N.S., not significant). (B) Western blotting shows FGFR3 expression levels of parental cells and cells treated with sicontrol, EGFP, siFGFR3, FGFR3IIIb, or FGFR3IIIc. β-actin was expressed equally among the cells. (C) RT-PCR shows that FGF2 was endogenously expressed in parental EC-GI-10 cells.

Article Snippet: The cell lysates were electrophoresed through 7.5% polyacrylamide gels and transferred to PVDF membranes (Immobilon-P; Millipore), and were analyzed by immunoblotting with the monoclonal anti-β-actin antibody (Sigma-Aldrich; 1:2000 dilution), anti-FGFR3 antibody (C-15) (Santa Cruz Biotechnology, Dallas, TX; 1:2000 dilution) and monoclonal anti-FGFR3IIIc antibody (cat#MAB7662, lot.GHK411041, R&D Systems; 1:500 dilution).

Techniques: Expressing, Over Expression, Infection, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: Journal of Histochemistry and Cytochemistry

Article Title: Enhanced Expression of Fibroblast Growth Factor Receptor 3 IIIc Promotes Human Esophageal Carcinoma Cell Proliferation

doi: 10.1369/0022155415616161

Figure Lengend Snippet: FGFR3IIIb and FGFR3IIIc Expression in Esophageal Carcinoma (EC) and Non-Cancerous Mucosa (NCM).

Article Snippet: The cell lysates were electrophoresed through 7.5% polyacrylamide gels and transferred to PVDF membranes (Immobilon-P; Millipore), and were analyzed by immunoblotting with the monoclonal anti-β-actin antibody (Sigma-Aldrich; 1:2000 dilution), anti-FGFR3 antibody (C-15) (Santa Cruz Biotechnology, Dallas, TX; 1:2000 dilution) and monoclonal anti-FGFR3IIIc antibody (cat#MAB7662, lot.GHK411041, R&D Systems; 1:500 dilution).

Techniques: Expressing

Journal: Journal of Histochemistry and Cytochemistry

Article Title: Enhanced Expression of Fibroblast Growth Factor Receptor 3 IIIc Promotes Human Esophageal Carcinoma Cell Proliferation

doi: 10.1369/0022155415616161

Figure Lengend Snippet: Gene expression of FGFR3 isoforms in esophageal carcinoma (EC) and non-cancerous mucosa (NCM). Total RNA was extracted from the specimens of EC patients. The gene expression of FGFR3 in 16 patients (E1 to E16) was analyzed by RT-PCR. FGFR3IIIc expression was clearly detected in the EC (T; tumor) of 12 patients (E2, E3, E4, E5, E6, E7, E8, E10, E13, E14, E15, and E16) and in the NCM (N; non-tumor) of 6 patients (E1, E3, E10, E13, E14, and E15). FGFR3IIIb expression was clearly detected in the EC (T) of 12 patients and in the NCM (N) of 11 patients. GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: The cell lysates were electrophoresed through 7.5% polyacrylamide gels and transferred to PVDF membranes (Immobilon-P; Millipore), and were analyzed by immunoblotting with the monoclonal anti-β-actin antibody (Sigma-Aldrich; 1:2000 dilution), anti-FGFR3 antibody (C-15) (Santa Cruz Biotechnology, Dallas, TX; 1:2000 dilution) and monoclonal anti-FGFR3IIIc antibody (cat#MAB7662, lot.GHK411041, R&D Systems; 1:500 dilution).

Techniques: Gene Expression, Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Calcified Tissue International

Article Title: Fibroblast Growth Factor 23 (FGF23) and Alpha-Klotho Stimulate Osteoblastic MC3T3.E1 Cell Proliferation and Inhibit Mineralization

doi: 10.1007/s00223-011-9501-5

Figure Lengend Snippet: FGFRs were expressed and modulated by FGF23 + Klotho, but not Klotho or FGF23 alone (B-DNA analysis). Cells were cultured in 12-well plates in differentiation medium (alpha-MEM/10% FBS/ascorbic acid [50 μg/mL]/BGP [10 mM]) and supplemented with huFGF23R176Q (F) (1 and 1,000 ng/mL), murine Klotho (KL) (50 ng/mL), their combination, or FGF2 (bFGF) for 14 days. a FGFR “IIIc” forms are preferentially expressed in MC3T3.E1 cells exposed to differentiation medium alone. b FGFR1(IIIc). c FGFR2(IIIc). d FGFR3(IIIc). Gene panel FGFR1(IIIb), FGFR1(IIIc), FGFR2(IIIb), FGFR2(IIIc), FGFR3(IIIc) and FGFR4. Bars represent mean ± SEM. * P < 0.05, ** P < 0.01 compared with differentiation medium ( Diff. Medium )

Article Snippet: The p38 inhibitor SB203580 (PHZ1253, lot 72547179A, stock 50 mg/mL, 132 mM) was from Invitrogen; the phosphoinositide 3-kinase (P13 K) inhibitor LY294002 (9901, lot 9, stock 15.3 mg/mL, 50 mM) was from Cell Signaling (Hayward, CA); FGFR2(IIIc) neutralizing antibody (MAB716, lot FSQ02, stock 5 mg/mL, 33 μM (ND50 = 0.67–2.7 nM) and FGFR3 neutralizing antibody (MAB710, lot FTD02, stock 5 mg/mL, 33 μM (ND50 = 20–80 nM) were from R&D Systems.

Techniques: Cell Culture