Acute Inducible Ablation of GRP78 Reveals Its Role in Hematopoietic Stem Cell Survival, Lymphogenesis and Regulation of Stress Signaling
Figure 1
GRP78 deficiency leads to lymphopenia.
A: GRP78 expression in WT BM subpopulations. (Left) Grp78 mRNA expression in WT BM subpopulations measured by quantitative real-time PCR. The experiments were performed in duplicates; each replicate contains pooled BM from two WT mice. (Right) GRP78 expression in LSKCD34− and LSKCD34+ subpopulations in WT mice (n = 4) measured by flow cytometry. The bar graph represents the medium intensities of GRP78 staining with LSK cells set as 1. B: (Upper) Representative PCR genotyping results from 78f/f and c78f/f BM 6 days post completion of pI.pC treatment. (Lower) Western blot results for detection of GRP78 protein level in the BM performed in duplicates. C: Organ size and morphology from mice of the indicated genotypes. Arrows on top of the heart indicate thymus. D: Quantitation of the thymus cellularity (n = 4 for 78f/f, n = 4 for c78f/f) and spleen weight (n = 14 for 78f/f, n = 20 for c78f/f). E: H&E staining of paraffin sections of thymus and spleen of 78f/f and c78f/f mice. Arrows indicate megakaryocytes in the spleen. The scale bar represents 200 µm in thymus and 20 µm in spleen. F: Peripheral lymphocyte count using complete blood count analysis with tail peripheral blood from (Left) 78f/f (n = 12), c78f/f (n = 16) mice and (Right) 78f/f(t) (n = 3), c78f/f(t) (n = 3) chimeric mice. All data are presented as mean ± s.e (**P<0.01, ***P<0.001, Student’s t test).