Qualitative rather than quantitative differences in the UPR on the B10 and NOD
backgrounds. (a) Globa principal-components analysis (PCA) of
RNA-seq data for islets (n = 3 mice/group).
(b) Schematic of the number of significant (corrected P
< 0.05) gene expression differences for pairs of sample
conditions. (c) Number of significant gene set changes in gene set
enrichment analysis (GSEA) using cutoffs of P < 0.05
and P < 0.001. (d) ERAI mice were crossed
to B10k.Rag1−/−.insHEL
and NODk.Rag1−/−.insHEL
mice, and islets were analyzed by flow cytometry. Histograms show Xbp1s (Venus)
expression in insulin-expressing beta cells. Results are representative of three
experiments. (e) PCA of the Xbp1 response gene set in islets
(n = 3 mice/group). (f) Scatterplot of
individual Xbp1 response genes, showing the average log2-transformed fold change
in expression induced by insHEL on the B10 background versus the NOD background.
The dashed line indicates equivalent regulation; outlier genes are annotated.
(g) Dot plot of mass spectrometry expression ratios, showing on
each axis the log2-transformed ratio of expression in
B10k.Rag1−/−.insHEL
islets as compared to
NODk.Rag1−/−.insHEL
islets in duplicate experiments. All reproducibly detected proteins are
displayed in gray, with black dots indicating annotated UPR-related proteins.
(h,i) Immunofluorescence of wild-type and insHEL transgenic
islets with a polyclonal antibody to insulin, antibody to Manf and DAPI.
Staining is representative of three experiments (h), with Manf
quantification (n = 10 mice/group) (i).
Scale bar, 100 μm. Data are shown as means ± s.e.m.
*P < 0.05,
***P < 0.0001. NS, not
significant.