Journal: Molecular Cancer

Article Title: Extranuclear ERα is associated with regression of T47D PKCα-overexpressing, tamoxifen-resistant breast cancer

doi: 10.1186/1476-4598-12-34

Figure Lengend Snippet: ERα/caveolin-1 complex formation in response to E2, TAM and RAL treatment in T47D:A18/PKCα tumors. A . Representative western blot of co-IP experiments in T47D:A18/PKCα and T47D:A18/neo tumor extracts as detailed in materials and methods. B . Densitometric quantification of three co-IP experiments from three independent tumors for each group. Error bars represent SEM. *, P < 0.05 compared to all groups determined by one-way ANOVA followed by Bonferroni’s post-test.

Article Snippet: The following antibodies were used: rabbit monoclonal ERα (for tissue and cells, SP1, Lab Vision, Thermo Scientific, Kalamazoo, MI USA), mouse monoclonal ERα (alternative epitope to confirm specificity for tissue, 1D5, N-terminal epitope, Abcam, Cambridge, MA USA), rabbit polyclonal ERα (for colonies, HC20, Santa Cruz Biotechnology, Santa Cruz, CA USA), and mouse monoclonal caveolin-1 (Clone2234, BD Transduction Laboratories, Franklin Lakes, NJ USA).

Techniques: Western Blot, Co-Immunoprecipitation Assay

Journal: Journal of Korean Medical Science

Article Title: Impact of Caveolin-1 Expression on the Prognosis of Transitional Cell Carcinoma of the Upper Urinary Tract

doi: 10.3346/jkms.2008.23.2.296

Figure Lengend Snippet: Immunohistochemical staining for caveolin-1. (A) Cancer cells were not stained (caveolin-1 intensity 0). (B) Smooth muscle cells were stained as an internal control. Cancer cells were weakly stained, compared to an internal control (caveolin-1 intensity 1). (C) Cancer cells were stained in same intensity as internal control (caveolin-1 intensity 2). (D) Cytoplasms of cancer cells were strongly stained for caveolin-1 (caveolin-1 intensity 3). Original magnification, ×400.

Article Snippet: Briefly, paraffin-embedded 4- m tissue sections were deparaffinized, treated with 3% hydrogen peroxide, followed by incubation with the blocking reagent, and then incubated with mouse monoclonal antibody against caveolin-1 (clone 2297, diluted 1:200; BD Biosciences, San Diego, CA, U.S.A.) for 1 hr at 37℃.

Techniques: Immunohistochemical staining, Staining

Journal: Journal of Korean Medical Science

Article Title: Impact of Caveolin-1 Expression on the Prognosis of Transitional Cell Carcinoma of the Upper Urinary Tract

doi: 10.3346/jkms.2008.23.2.296

Figure Lengend Snippet: Kaplan-Meier cancer-specific survival curves according to the caveolin-1 expression. The survival rate of patients with caveolin-1 positive tumors was significantly lower than that of patients with caveolin-1 negative tumors ( p <0.0001).

Article Snippet: Briefly, paraffin-embedded 4- m tissue sections were deparaffinized, treated with 3% hydrogen peroxide, followed by incubation with the blocking reagent, and then incubated with mouse monoclonal antibody against caveolin-1 (clone 2297, diluted 1:200; BD Biosciences, San Diego, CA, U.S.A.) for 1 hr at 37℃.

Techniques: Expressing

Journal: Journal of Biomedical Materials Research. Part B, Applied Biomaterials

Article Title: Safety of intradiscal injection and biocompatibility of polyester amide microspheres in a canine model predisposed to intervertebral disc degeneration

doi: 10.1002/jbm.b.33579

Figure Lengend Snippet: A–G. Relative gene expression levels in non, sham‐, and PEAM‐injected, canine IVDs at 6 months follow‐up. The noninjected nuclei pulposi (NP) are set at 1. A–C. Gene expression levels of COL2A1 ( A ) and COL1A1 ( B ) were not significantly different between conditions, neither was collagen corrected for DNA ( C ). D. CASP3 expression levels were significantly lower in the AF compared with the NP. E. Expression levels of CAV1 were significantly higher in sham‐injected IVDs (NP + AF) compared with noninjected control IVDs. F. In all conditions glycosaminoglycan (GAG) corrected for DNA were significantly higher in the NP compared with the AF. G. The PEAM‐injected IVDs showed a significant higher BAX/BCL2 ratio compared with the sham‐injected IVDs. Data in A , B , D , and E are expressed as n ‐fold changes, in C and F as mean values, and in G as ratio ± standard deviation. * Indicate significant difference at a 99% confidence level

Article Snippet: Paraffin sections (5 µm) stained with hematoxylin/eosin and with picrosirius red/alcian blue were histopathologically evaluated by two independent investigators, blinded to the treatments, according to the grading scheme developed by Bergknut et al. Immunohistochemistry for caveolin‐1 [monoclonal mouse anti‐caveolin‐1 antibody (Clone 2297, 610406, BD Biosciences) diluted 1:50 in PBS] was performed as described previously.

Techniques: Expressing, Injection, Standard Deviation

Journal: Investigative and Clinical Urology

Article Title: Decreased urothelial expression of caveolin 1 and 2 in aging rats showing detrusor overactivity: Potential association with aging bladder

doi: 10.4111/icu.20210284

Figure Lengend Snippet: Immunofluorescence labeling for caveolin 1 and caveolin 2. Distinct staining of caveolin 1 and 2 was apparent subepithelial area in the urothelium (→ urothelium, *suburothelium). Immunofluorescence staining showed that the old-aged group had decreased expression of caveolin 1 (p=0.01) and caveolin 2 (p=0.02). The horizontal scale bar at the bottom left of each figure indicates the magnification power. The right panels denote the mean±standard deviation of 5 experiments for each condition determined by relative densitometry. * p<0.05.

Article Snippet: Monoclonal mouse antibodies for caveolin 1 and caveolin 2 (1:1,000; BD Biosciences) and polyclonal rabbit antibody against glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:4,000; Cell Signaling Technology, Danvers, MA, USA) were used.

Techniques: Immunofluorescence, Labeling, Staining, Expressing, Standard Deviation

Journal: Investigative and Clinical Urology

Article Title: Decreased urothelial expression of caveolin 1 and 2 in aging rats showing detrusor overactivity: Potential association with aging bladder

doi: 10.4111/icu.20210284

Figure Lengend Snippet: Immunoblotting of caveolin 1 and caveolin 2. Western blot analysis revealed bands at 20 kDa corresponding to caveolin 1 and caveolin 2 proteins. Anti-GAPDH antibody recognized the 42 kDa band. The protein expression of both caveolin 1 and 2 was significantly decreased in the old-aged group than in the young age control group. The lower panels denote the mean±standard deviation of 5 experiments for each condition determined by densitometry relative to GAPDH. * p<0.05.

Article Snippet: Monoclonal mouse antibodies for caveolin 1 and caveolin 2 (1:1,000; BD Biosciences) and polyclonal rabbit antibody against glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:4,000; Cell Signaling Technology, Danvers, MA, USA) were used.

Techniques: Western Blot, Expressing, Standard Deviation

Journal: The Journal of Biological Chemistry

Article Title: S -Palmitoylation of junctophilin-2 is critical for its role in tethering the sarcoplasmic reticulum to the plasma membrane

doi: 10.1074/jbc.RA118.006772

Figure Lengend Snippet: JPH2 in lipid–raft domain is enriched with palmitoylated form, and inhibiting JPH2 palmitoylation reduces its presence in the lipid–raft domains. A, raft and nonraft components of cell membranes from COS-7 cells expressing FLAG–JPH2 labeled with palmitate–alkyne were separated by detergent-free purification/sucrose-gradient fractionation. Fractions were analyzed by immunoblots (IB) with JPH2 and caveolin-1 (Cav-1, raft marker) Abs. Fractions 6 and 7 with high Cav-1 contents were combined as “Raft” component, and fractions 11 and 12 with the lowest Cav-1 contents were combined as “Non-raft” component. Both components were solubilized, and FLAG–JPH2 was immunoprecipitated and reacted with biotin–PEG3–azide in the CuAAC reaction. B, immunoblot images of CuAAC reacted IPs from raft and nonraft components probed with JPH2 Ab and then biotin Ab. C, degree of JPH2 palmitoylation was quantified by dividing the biotinylation band intensity (100 kDa) by the total JPH2 band intensity at 100 kDa. D, Triton-soluble and -insoluble fractions (Sol and Insol) were isolated from COS-7 cells expressing FLAG–JPH2 (without or with 2BP pretreatment), WT, and Cys-free JPH2–GFP or mCherry–JPH2, and analyzed by immunoblot with JPH2, Cav-1, and α-actin Abs (the latter as loading control). FLAG–JPH2 migrated as 100- and 75-kDa bands in Sol lanes (closed and open circles), but as a single 100-kDa band in Insol lanes. JPH2–GFP and mCherry–JPH2 migrated as 125- and 100-kDa bands in Sol lanes, but as single 125 kDa in Insol lanes. Cys-free JPH2–GFP migrated as a single 100-kDa band in the Sol lane and was barely detectable in the Insol lane. Image of the right six lanes from the same membrane is shown in high contrast to better illustrate the differential banding pattern described above. E, bar plot of distribution of JPH2 variants between Insol and Sol components, estimated by dividing the JPH2 band intensity in the Insol lane by the combined JPH2 band intensities in the Sol lane.

Article Snippet: The following primary antibodies were used: JPH2 mouse and rabbit Abs (Santa Cruz Biotechnology, sc-377086 and sc-134875); mCherry rabbit Ab (Abcam, ab167453); GFP rabbit Ab (Abcam, ab290); FLAG mouse Ab (Sigma, F3165); dsRed rabbit Ab (Clontech, 632496); caveolin-1 and caveolin-3 mouse Abs (BD Transduction Laboratories, 610057 and 610421); biotin goat Ab (Sigma, B3640); Cav1.2 mouse Ab (NeuroMab, 75–257); and α-actin mouse Ab (Sigma, A7811).

Techniques: Expressing, Labeling, Purification, Fractionation, Western Blot, Marker, Immunoprecipitation, Isolation