Journal: International Journal of Molecular Sciences

Article Title: Differentiated PDGFRα-Positive Cells: A Novel In-Vitro Model for Functional Studies of Neuronal Nitric Oxide Synthase

doi: 10.3390/ijms22073514

Figure Lengend Snippet: Expression of CD140α, CD44, CD34 and SK3 cell surface markers in iMSCs, fibroblasts, and PDGFRα-positive cells. Flow cytometry analysis detected the expression of CD140α (( A ). cell count; ( B) . mean fluorescence intensity), CD44 (( C ). cell count; ( D ). mean fluorescence intensity), CD34 (( E ). cell count; ( F ). mean fluorescence intensity), and SK3 (( G ). cell count; ( H ). mean fluorescence intensity) in iMSCs, fibroblasts, and PDGFRα-positive ( n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001); ns: not significant.

Article Snippet: Cells were then washed with PBS for 10 min and incubated with the following primary antibodies in a humidified chamber overnight at 4 °C: anti-CD34 (ab81289; Abcam, Cambridge, UK), anti-CD44 (BD Pharmingen, San Diego, CA, USA), anti-KCa2.3-ATTO-594 (SK3; Alomone Labs, Jerusalem, Israel), anti-PDGFRα (ab234965; Abcam, Cambridge, UK), and anti-nNOS (3G6B10; Invitrogen Carlsbad, CA, USA).

Techniques: Expressing, Flow Cytometry, Cell Counting, Fluorescence

Journal: American Journal of Physiology - Renal Physiology

Article Title: Caveolae facilitate TRPV4-mediated Ca 2+ signaling and the hierarchical activation of Ca 2+ -activated K + channels in K + -secreting renal collecting duct cells

doi: 10.1152/ajprenal.00076.2018

Figure Lengend Snippet: Representative immunofluorescence images show colocalization of transient receptor potential cation channel subfamily V member 4 (TRPV4) with each of the KCa channels in collecting duct mCCDcl1 cells in mouse kidney cortical collecting duct (CCD). mCCDcl1 cells immunostained for TRPV4 (TRPV4, green) and SK3 (SK3-ATTO-594, red) show strong colocalization as apparent in the merged image (Merged, yellow) (A). Similarly, immunostaining for TRPV4 (red) and BKα (green) (B) and for TRPV4 (red) and IK1 (green) (C) demonstrated noted colocalization in the merged images (Merged, yellow). Mouse kidney CCD immunostained for TRPV4 (TRPV4, green) and SK3 (SK3, ATTO-594, red) display noted colocalization in the merged images (Merged, yellow) (D). In a similar manner, immunostaining for TRPV4 (TRPV4, red) and BKα (BKα, green) (E) and for TRPV4 (TRPV4, red) and IK1 (IK1, green) (F) likewise demonstrated apparent colocalization in the merged images (Merged, yellow). Areas of more diffuse staining are also apparent in some cases, especially for IK1 (see F). Note that we verified the primary antibodies using blocking peptides in an earlier publication [(20) Figs. 1 and ​and4].4]. All the immunolocalization experiments were repeated at least three times (n = 3). BK, large conductance Ca2+-activated K+ channel; IK, intermediate conductance Ca2+-activated K+ channel; SK, small conductance Ca2+-activated K+ channel.

Article Snippet: The following primary antibodies were used: anti-CAV-1 (Thermo Fisher, cat. no. MA-3-600), anti-SK1 (Alomone, cat. no. APC-039), anti-SK3-ATTO-594 (Alomone, cat. no. APC-025-AR), anti-IK1 (Alomone, cat. no. ALM-051), anti-BKα (Alomone, cat. no. APC-151), and anti-TRPV4 (Alomone, cat. nos.

Techniques: Immunofluorescence, Immunostaining, Staining, Blocking Assay