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Table 1.

Active PKC isoforms detected in undifferentiated and differentiated CACO-2 cultures

Protein P-site tested 4 Days 8 Days % Increase % Decrease Polarity Ref.
PKC iota (ι)   T555   4985   6613   33    Apical   (Suzuki and Ohno, 2006)  
PKC alpha (α)   S657   156   482   309    Apical   (Song et al., 2001)  
PKC beta (β)   T641   340   190    45   Basolateral   (Padanilam, 2001)  
PKC delta (δ)   S645   683   402    42   Basolateral   (Song et al., 2001)  
PKC epsilon (ϵ)   S729   31   30     Basolateral   (Song et al., 2001)  
PKC eta (η)   T655   Not detected   Not detected      
PKC gamma (γ)   T514   135   419   310*    
PKC theta (θ)   S676   Not detected   Not detected    Tight junction   
PKC theta (θ)   S695   Not detected   Not detected      
PKC theta (θ)   T538   Not detected   Not detected      
PKC zeta (ζ)   T410/403   534   545     Apical   (Padanilam, 2001)  
Protein P-site tested 4 Days 8 Days % Increase % Decrease Polarity Ref.
PKC iota (ι)   T555   4985   6613   33    Apical   (Suzuki and Ohno, 2006)  
PKC alpha (α)   S657   156   482   309    Apical   (Song et al., 2001)  
PKC beta (β)   T641   340   190    45   Basolateral   (Padanilam, 2001)  
PKC delta (δ)   S645   683   402    42   Basolateral   (Song et al., 2001)  
PKC epsilon (ϵ)   S729   31   30     Basolateral   (Song et al., 2001)  
PKC eta (η)   T655   Not detected   Not detected      
PKC gamma (γ)   T514   135   419   310*    
PKC theta (θ)   S676   Not detected   Not detected    Tight junction   
PKC theta (θ)   S695   Not detected   Not detected      
PKC theta (θ)   T538   Not detected   Not detected      
PKC zeta (ζ)   T410/403   534   545     Apical   (Padanilam, 2001)  

Identical amounts of protein from confluent CACO-2 cells at 4 days (undifferentiated) or 8 days (differentiated) after seeding were analyzed by immunoblot using a pre-established panel of specific anti-phospho-epitope antibodies that recognize the active form of each kinase. The facility performing the analysis (Kinexus, Vancouver, Canada) has normalized the arbitrary units, so that the results are comparable, not only for each kinase, but also among different kinases.

*

PKC gamma is normally negative in the intestine and characteristic of neural tissue (Jiang et al.,1995), although present in some colon cancers in low levels which explains its presence in CACO-2 cells (Doi et al., 1994).

Although we did not detect it with three different antibodies, Keshavarzian and coworkers (Banan et al., 2004) recently reported a small amount of PKCθ in CACO-2 cells, but colocalizing with claudins at the tight junctions.

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