Gene expression analysis in lacrimal glands.

<p>A. Tumor necrosis factor-α (TNF-α). p = 0.06, N = 10, average ± standard error of mean (SE). B. Monocyte chemotactic protein-1 (MCP-1). *p = 0.01, N = 10, average ± SE. MCP-1 and TNF-α expression in the lactoferrin (LF) group were lower than the control (CTL) group.</p

Tear secretion volume.

<p>A. Time course of the tear secretion. B. Lacrimal gland weight. C. Tear secretion corrected by lacrimal gland weight at 6 months. The volume of tear secretion was significantly larger in the lactoferrin (LF) group compared to the control (CTL) group. *p<0.01, N = 10, average ± standard error of mean).</p

Body weight and daily food intake.

<p>Body weight and daily food intake.</p

Oxidative stress markers in serum.

<p>A. 8-hydroxy-2′-deoxyguanosine (8-OHdG). *p = 0.004, N = 5, average ± standard error of mean (SE). B. Hexanoyl-lysine adduct (HEL). p = 0.06, N = 5, average ± SE. The concentration of 8-OHdG and HEL in the lactoferrin (LF) group were lower than the control (CTL) group.</p

Primers.

<p>Primers.</p

Morphological changes in lacrimal glands.

<p>Hematoxylin-eosin staining of control (A, C) and lactoferrin groups (B, D). CD45 immunostaining of lacrimal glands in the control group (E) and the lactoferrin group (F) Organelle morphology of control (G) and lactoferrin groups (H) examined by transmission electron microscope. Scale bar indicated 100 µm (A–F) and 2 µm (G, H).</p

Serum parameters.

*<p>p<0.05.</p

Gene expression analysis in conjunctivae.

<p>A. TNF-α. p = 0.36, N = 7, average ± standard error of mean (SE). B. MCP-1. p = 0.06, N = 7, average ± SE. MCP-1 and TNF-α expression in the lactoferrin (LF) group tended to belower than the control (CTL) group.</p

Oxidative stress marker in lacrimal glands.

<p>8-OHdG immunostaining of lacrimal glands in the control group (A) and the lactoferrin group (B). Scale bar: 50 µm.</p

Infiltration of inflammatory mononuclear cells in <i>Itpr2^−/−</i>;<i>Itpr3^−/−</i> lacrimal glands.

<p>(A) Immunostaining of CD45, F4/80, CD19, CD8 and CD4 in lacrimal gland tissue sections from wild-type and <i>Itpr2^−/−</i>;<i>Itpr3^−/−</i> mice. White arrowheads indicate inflammatory mononuclear cells. (B) Quantification of TNF-α mRNA expression levels by real time RT-PCR. Six week-old mice; wild-type: n = 8 and <i>Itpr2^−/−</i>;<i>Itpr3^−/−</i>: n = 8. Ten week-old mice; wild-type: n = 16, <i>Itpr2^−/−</i>;<i>Itpr3^−/−</i>: n = 10. Mann–Whitney <i>U</i>-test, ***P<0.001, *P<0.05. All data are presented as means ± SEM. (C) Quantification of IL-6 mRNA expression levels by real time RT-PCR. Six week-old mice; wild-type: n = 8 and <i>Itpr2^−/−</i>;<i>Itpr3^−/−</i>: n = 8. Ten week-old mice; wild-type: n = 16, <i>Itpr2^−/−</i>;<i>Itpr3^−/−</i>: n = 10. Mann–Whitney <i>U</i>-test, ***P<0.001. All data are presented as means ± SEM.</p