Figure 3.

Apical vs basolateral TNF-α signaling. (A) Monolayers grown in 96-well transwells (5 wells per condition) were challenged with an 8-point dose curve of TNF-α on either the apical or basolateral sides of the monolayer. TEER measurements were taken at 72-h post-challenge and analyzed in a similar manner to Fig. 2B. (B) Quantifying differences in transcriptional response to 40 ng/mL TNF-α challenge on either side of monolayers. 6 h post-challenge, monolayers grown in 24-well transwells (3 wells per condition) were lysed and processed for qPCR. Gene expression for IL1B and TNF were quantified, and fold change values were calculated by normalizing to untreated controls. (C) Quantifying the potency of anti-TNF-α neutralizing antibodies by different routes of delivery. Monolayers grown in 96-well transwells (5 wells per condition) were pre-treated with anti-TNF-α antibody on either apical or basolateral sides before TNF-α challenge on the basolateral side. TEER measurement was taken 72 h post-challenge and EC50 values and curve fit was performed in the same manner as Fig. 2C.