rabbit anti-snap23  (Thermo Fisher)

Journal: Cell Communication and Signaling : CCS

Article Title: TGF-β2 increases eHSP90α secretion via upregulating secretory autophagy pathway

doi: 10.1186/s12964-025-02493-5

Figure Lengend Snippet: Knockdown of Q-SNARE STX3, STX4 and SNAP23 inhibits eHSP90α secretion. A Immunoblotting HSP90α and STX3 in supernatants and cell lysates of SRA01/04 that were transfected with scramble siRNA or siRNA against STX3 followed by treatment with 10 ng/ml TGF-β2 for 4 h. B Quantitation of eHSP90α vs. total HSP90α in Fig. 5A by Image J. C Immunoblotting HSP90α and STX4 in supernatants and cell lysates of SRA01/04 that were transfected with scramble siRNA or siRNA against STX4 followed by treatment with 10 ng/ml TGF-β2 for 4 h. D Quantitation of eHSP90α vs. total HSP90α in Fig. 5C. E Immunoblotting HSP90α and SNAP23 in supernatants or cell lysates of SRA01/04 that were transfected with scramble siRNA or siRNA against SNAP23 followed by treatment with TGF-β2 for 4 h. F Quantitation of eHSP90α vs. total HSP90α in Fig. 5E. All the data shown are mean ± SD, n = 3. One-way ANOVA statistical analysis for Fig. 5B, D and F. *** p < 0.01. N.S., no significance

Article Snippet: Rabbit anti-ATG7 (A19604), anti-SEC22B (A15358), anti-STX3 (A3712), anti-STX4 (A5996), SNAP23 (A13909), anti-pan Akt (A18675) and anti-p38 (A14401) antibodies were from Abclonal Technology (Wuhan, China).

Techniques: Knockdown, Western Blot, Transfection, Quantitation Assay

Journal: bioRxiv

Article Title: Porosome reconstitution therapy: A biologic rescue from cystic fibrosis

doi: 10.1101/2024.09.11.612494

Figure Lengend Snippet: (A) Wide-field fluorescence images of untreated HBE cells (left) and HBE cells with CFTR ΔF508 mutation treated with increasing concentrations of porosomes (0.5, and 1 µg). The scale bar represents 50 µm. DAPI (blue) stains the nuclei, and CFTR (green) highlights the CFTR protein. ( B) Magnify-processed images of the same conditions as in Panel A, showing enhanced resolution and detail of CFTR expression. The scale bar represents 50 µm. ( C) Zoomed-in images of the areas indicated by yellow-dotted boxes in Panel B. These images provide a closer view of CFTR expression in the HBE cells. The scale bar represents 5 µm. (D) Magnify-processed images of CFTR ΔF508-mutated HBE cells treated with increasing doses of porosomes (0.2, 0.5, and 1 µg). This panel shows DAPI (blue), CFTR (green), and SNAP23 (red), illustrating the expression and localization of both proteins in response to porosome treatment. The scale bar represents 5 µm in biological scale. B, C and D: The expansion factor is 3.8× (1× PBS).

Article Snippet: Cells were incubated with a staining buffer (2X SSC, 0.2% Tween-20, 1X PBS) containing primary antibodies (1:500 dilution): mouse anti-CFTR (Invitrogen; MA1-935) and rabbit anti-SNAP23 (Invitrogen; PA5-28936) for 45 minutes to 1 hour at room temperature.

Techniques: Fluorescence, Mutagenesis, Expressing

Journal: eLife

Article Title: ICAM-1 nanoclusters regulate hepatic epithelial cell polarity by leukocyte adhesion-independent control of apical actomyosin

doi: 10.7554/eLife.89261

Figure Lengend Snippet: ( A ) Schematic representation of the proximal biotinylation of proteins by the ICAM-1-BirA * chimeric protein. ( B ) Cells expressing ICAM-1-BirA* or not were incubated with 50 μM biotin for 16 hr, lysed and subjected to a pull-down (PD) assay with neutravidin-agarose. Biotinylated proteins were detected by blot with streptavidin-HRP. ( C ) PD fractions from two experiments were subjected to mass spectrometry analysis. Table shows the proteins identified exclusively in the fraction of HepG2 expressing ICAM-1-BirA * but not in the PD fraction of parental cells. EBP50 proximal interaction with ICAM-1 is validated by western blot in ( A ). ( D ) Proximal interaction of SNAP23, but not of ezrin-radixin-moesin proteins, despite both having been previously described as ICAM-1 interactors ( ; ). ( E ) Cells stably expressing the indicated Flag-tagged EBP50 proteins were cultured for 48 hr, fixed and stained for the FLAG epitope and for F-actin to localized bile canaliculi (BC). Scale bars, 5 μm. ( F ) Cells were incubated with anti-ICAM-1 mAb for 30 min at 4°C, washed and incubated at 37°C for the indicated times. Cells were lysed and soluble and insoluble (pellet) fractions were separated by centrifugation as described in ‘Materials and methods’. The relative distribution of the indicated proteins was analyzed by western blot (left) and quantified (right). Figure 6—figure supplement 1—source data 1. Original file for the western blot analysis in (neutravidin-HRP). Figure 6—figure supplement 1—source data 2. PDF containing and original scan of the relevant western blot (neutravidin-HRP), with the highlighted region squared. Figure 6—figure supplement 1—source data 3. Original file for the western blot analysis in (anti-ICAM-1 and anti-SNAP23). Figure 6—figure supplement 1—source data 4. Original file for the western blot analysis in (anti-ERMs). Figure 6—figure supplement 1—source data 5. PDF containing and original scan of the relevant western blot (anti-ICAM-1, anti-ERMs, and anti-SNAP23), with the highlighted region squared. Figure 6—figure supplement 1—source data 6. Original file for the western blot analysis in (anti-ICAM-1). Figure 6—figure supplement 1—source data 7. Original file for the western blot analysis in (anti-EBP50). Figure 6—figure supplement 1—source data 8. Original file for the western blot analysis in (anti-tubulin). Figure 6—figure supplement 1—source data 9. Original file for the western blot analysis in (anti-ERMs). Figure 6—figure supplement 1—source data 10. Original file for the western blot analysis in (anti-GAPDH). Figure 6—figure supplement 1—source data 11. PDF containing and original scan of the relevant western blot (anti-ICAM-1, anti-EBP50, anti-tubulin, anti-ERMs, and anti-GAPDH), with the highlighted region squared.

Article Snippet: Antibody , Anti-SNAP23 (rabbit polyclonal) , Synaptic Systems , 111 202; RRID: AB_887788 , WB 1/1000; IF 1/400.

Techniques: Expressing, Incubation, Mass Spectrometry, Western Blot, Stable Transfection, Cell Culture, Staining, FLAG-tag, Centrifugation

Journal: eLife

Article Title: ICAM-1 nanoclusters regulate hepatic epithelial cell polarity by leukocyte adhesion-independent control of apical actomyosin

doi: 10.7554/eLife.89261

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-SNAP23 (rabbit polyclonal) , Synaptic Systems , 111 202; RRID: AB_887788 , WB 1/1000; IF 1/400.

Techniques: Isolation, Transfection, Construct, Plasmid Preparation, CRISPR, Purification, Sequencing, Recombinant, Immunohistochemistry, Immunofluorescence, Cryo-EM Sample Prep, Tomography, Immunoprecipitation