Fig 2: B cell TNF and ADAM17 regulation in C57Bl/6 and Balb/c ADAM10B-/- (A10K0) and WT.(A) sTNF or (B) mTNF from 3 day stimulated (LPS + IL4) B cell cultures; (B) Balb/c WT (black), Balb/c A10KO (red), C57Bl/6 WT (green), and C57Bl/6 A10KO (blue) B cells. (C) Fold change (A10KO over WT) in relative TNF message from naïve (white) or 3 day stimulated (anti-CD40+IL4) (black) B cells normalized to 18s. (D) Fold change (A10KO over WT) in relative ADAM17 expression normalized to 18s for naïve (white) or 3 day stimulated (black) B cells. (C,D) * signifies ≥2 fold change between groups. (E) ADAM17 (~93kDa) and actin (~42kDa) from 5 day stimulated (anti-CD40 + IL4) B cells (left) and band densitometry (right). KO = A10KO. n = 9 per group, 3 independent studies. *p<0.05, **p<0.005.

Fig 3: Differences in the response of cultured SCG and PVG neurons to TNFR1‐activated TNF‐mediated reverse signaling. Sholl plots of P0 SCG (A) and PVG (B) neurons cultured for 18 h with either 10 ng/ml NGF or NGF plus 30 ng/ml TNFR1‐Fc. Mean ± s.e.m. of data of >150 neurons per condition combined from three separate experiments of each type. (C) Images of representative P0 SCG neurons and PVG neurons grown for 18 h with NGF with or without TNFR1‐Fc as indicated. Scale bar, 50 μm. Sholl plots of P0 SCG (D) and PVG (E) neurons cultured for 18 h with either 10 ng/ml NGF or NGF plus 50 ng/ml TNFR2‐Fc. Mean ± s.e.m. of data of >150 neurons per condition combined from three separate experiments of each type. [Colour figure can be viewed at wileyonlinelibrary.com]

Fig 4: Images of sympathetic innervation of the stomach and spleen of mice possessing null mutations in the Tnf and Tnfrsf1a genes. Representative images of P10 whole‐mount (A, C) and P6 iDISCO (B, D) preparations of the stomach and of P10 whole‐mount (E, G) and P6 iDISCO (F, H) preparations of the spleen of Tnf−/− and Tnf+/+ littermates (A, B, E, F) and Tnfrsf1a−/−and Tnfrsf1a+/+ littermates (C, D, G, H) stained with anti‐TH. Scale bars, 200 μm.

Fig 5: Immunolocalization of TNF, TNFR1, and TNFR2 on SCG and PVG neurons. Representative photomicrographs of P0 SCG neurons and PVG neurons double labeled with either anti‐TNFR1 and anti‐βIII‐tubulin antibodies (A) or anti‐TNF and anti‐βIII‐tubulin antibodies (B) or anti‐TNFR2 and anti‐βIII‐tubulin antibodies (C) after 24‐h culture in NGF‐supplemented medium. The preparations were also counterstained with the nuclear marker, DAPI. Scale bars, 10 μm. [Colour figure can be viewed at wileyonlinelibrary.com]