1. Volume 127, Issue 1, Pages 203-212 (July 2004)
Age and diet act through distinct isoforms of the class II transactivator gene in mouse intestinal epithelium 
Ian R. Sanderson, Stephen A. Bustin, Suzan Dziennis, Joanna Paraszczuk, Demetra S. Stamm 
Gastroenterology 
Volume 127, Issue 1, Pages (July 2004)
DOI: /j.gastro

2. Figure 1
Expression of CIITA depends on the transcription of 3 possible isoforms. The differences between them lie in the alternative start site on the CIITA gene and consequent distinct exons 1. Exon 2 and mRNA 3′ are common to all 3 isoforms of CIITA. Transcripts from the 3 different isoforms can be distinguished by PCR in which the 5′ primer hybridizes with sequences in exon 1, and the 3′ primer with those in exon 2.
Gastroenterology  , DOI: ( /j.gastro )

3. Figure 2
The standard curve shows the number of PCR cycles that were required to detect increasing standard concentrations of CIITA III and IV mRNA from intestinal epithelial cells of wild-type mice fed either elemental or complex diets (correlation coefficient for CIITA III = 0.98, and for CIITA IV = 0.99).
Gastroenterology  , DOI: ( /j.gastro )

4. Figure 3
A complex diet induces the transcription of Ii chain. Mice from a single litter were weaned on day 18 onto either an elemental diet (Vivonex) or Vivonex with 4 g of casein per 100 mL. RNA was isolated from mice killed in pairs up to day 40. Ii chain mRNA and β-actin was identified by using Northern blots. Data are representative of 3 experiments with similar results.
Gastroenterology  , DOI: ( /j.gastro )

5. Figure 4
Invariant chain mRNA is expressed by day 31 in the intestinal epithelium of mice weaned onto a complex diet but not in mice weaned onto an elemental diet. Furthermore, CIITA expression is induced in mice fed a complex diet but not those fed an elemental diet. RNase protection assay (CIITA mRNA) and Northern blot analysis (Ii chain mRNA) of RNA from epithelial cells taken from individual wild-type (C57BL/6) mice of a single split litter weaned at 18 days onto either an elemental or a complex (chow) diet. Samples were also probed by using β-actin to verify equal amounts of sample RNA. Data are representative of 4 litters.
Gastroenterology  , DOI: ( /j.gastro )

6. Figure 5
CIITA IV mRNA increases in the intestinal epithelium by day 31 onto a complex diet, but CIITA III does not. The Figure shows mRNA extracted at 2 experimental time points (days 24 and 31, as in Figure 4). Bars represent units of mRNA as a proportion of the mRNA measured on day 24 in mice weaned to an elemental diet (mean and standard deviation). CIITA IV mRNA increased postweaning onto normal complex (chow) diet (P < 0.02, n = 3). Results are representative of 3 separate experiments.
Gastroenterology  , DOI: ( /j.gastro )

7. Figure 6
IFN-γ induces transcription of CIITA IV (but not CIITA III) in mice weaned onto an elemental diet. Littermates were weaned on day 18 onto an elemental diet (n = 4) or onto normal chow diet (n = 2). Two mice on the elemental diet were given 100,000 units of IFN-γ on day 43 and again on day 44. RNA from intestinal epithelial cells was isolated on day 45 for all 6 mice. There were no significant differences in CIITA III expression. However, CIITA IV was significantly increased by IFN-γ (P < 0.02) and by weaning onto mouse chow (P < 0.02). Bars represent units of mRNA relative to that observed in elemental diet fed mice without IFN-γ (mean and standard deviation).
Gastroenterology  , DOI: ( /j.gastro )

8. Figure 7
Three-month-old IFN-γ receptor-deficient mice express class II MHC (Ia) on the intestinal epithelium (×60). (A) Sections were stained with rat anti-mouse I-A/I-E (M5/ ) antibody and biotinylated anti-rat immunoglobulin stained with diaminobenzidine. (B) Control sections were concurrently examined without primary antibody.
Gastroenterology  , DOI: ( /j.gastro )

9. Figure 8
Northern blot analysis shows the accumulation of Ii mRNA in the intestinal epithelium at different ages in wild-type and IFN-γ receptor knockout mice weaned onto elemental or complex (normal) diets. Wild-type mice weaned to a complex diet fully express Ii by day 31, whereas wild-type mice weaned onto an elemental diet only partially express Ii by day 45. At day 45, IFN-γ receptor knockout mice express a small amount of Ii chain mRNA irrespective of diet. Data are representative of 3 separate experiments.
Gastroenterology  , DOI: ( /j.gastro )

10. Figure 9
CIITA IV mRNA levels increase in the intestinal epithelium at day 45 in mice fed a complex diet. Bars represent units of mRNA as a proportion of the mRNA measured on day 24 in mice weaned to an elemental diet (mean and standard deviation). Neither wild-type nor IFN-γ receptor knockout mice fed elemental diets show any significant difference in CIITA IV mRNA levels at day 24 and day 45 postweaning (P = 0.3 and P = 0.2, respectively). However, there were significant differences between CIITA IV mRNA levels at the same time points in wild-type mice fed a complex diet (P < 0.05). The CIITA IV in chow-fed wild-type mice was significantly greater than in chow-fed IFN-γ receptor knockout mice (P < 0.01).
Gastroenterology  , DOI: ( /j.gastro )

11. Figure 10
CIITA III mRNA levels increase with time, irrespective of diet (samples as in Figure 8). Bars represent units of mRNA, expressed as a proportion of the mRNA measured on day 24 in mice weaned onto an elemental diet (mean and standard deviation). There are significant differences in CIITA III mRNA levels at day 24 compared with day 45 in both wild-type and IFN-γ receptor knockout mice fed elemental diets (P < 0.03) and also in both groups of mice fed complex diets (P < 0.01 and P < 0.04, respectively).
Gastroenterology  , DOI: ( /j.gastro )

12. Figure 11
CIITA III increases with time, despite switching from a complex (chow) diet to an elemental diet. Mice (n = 8) were weaned onto mouse chow at day 18. mRNA was isolated from intestinal epithelium from 2 mice on day 31. The remainder were randomized either to continue chow diet (n = 3) or were changed to an elemental diet on day 31 (n = 3). RNA was isolated from the intestinal epithelium of both groups on day 45. There was a significant increase in the CIITA III between day 31 and day 45, irrespective of whether mice remained on chow (P < 0.02) or were changed onto an elemental diet (P < 0.02). There was no significant change in CIITA IV in mice whose diet was switched from chow to an elemental diet. In this Figure, CIITA is measured relative to that of postweaned mice on day 31 and not to preweaned mice as in other Figures.
Gastroenterology  , DOI: ( /j.gastro )