Extended Data Fig. 2: PIKfyve is critical for progression of precursor PanIN lesions to PDAC.
From: Targeting PIKfyve-driven lipid metabolism in pancreatic cancer
a. Representative images of PIKfyve BaseScope staining from pancreas tissue of 27-week-old KC Pikfyve+/+, KC Pikfyvef/+, and KC Pikfyvef/f mice. Scalebar = 60 μm for zoomed-out images; 30 μm for zoomed-in images. b. Pikfyve levels as determined by BaseScope of KC Pikfyve+/+, KC Pikfyvef/+, and KC Pikfyvef/f murine pancreas tissue separated by normal and lesional areas. Box represents 25th and 75th percentiles; whiskers represent the range. (One way ANOVA with Dunnett’s.). c. Pancreas tissue weight from 27-week-old KC Pikfyve+/+, KC Pikfyvef/+, KC Pikfyvef/f, and age-matched wild-type (WT) mice. Box represents 25th and 75th percentiles; whiskers represent the range. (One-way ANOVA with Dunnett’s.). d. Pancreas tissue weight normalized to total body weight from KC Pikfyve+/+, KC Pikfyvef/+, KC Pikfyvef/f or age-matched wild-type (WT) mice at 27 weeks of age. Box represents 25th and 75th percentiles; whiskers represent the range. (One-way ANOVA with Dunnett’s.). e. Percentage of pancreas occupied by normal tissue as determined by histological analyses in KC Pikfyve+/+, KC Pikfyvef/+, and KC Pikfyvef/f mice at 27 weeks of age. Bars are +/- SEM. (One-way ANOVA with Dunnett’s.). f. Representative histological images showing H&E and CK19 staining on pancreatic tissue of KC Pikfyve+/+, KC Pikfyvef/+, and KC Pikfyvef/f mice at 27 weeks of age. Scalebar = 5 mm for the whole-pancreas images, 300 μm for the zoomed-in images. g. Raw pancreas tissue weight (top) and pancreatic weight normalized to total body weight (bottom) from 40-week-old KC Pikfyve+/+, KC Pikfyvef/+, and KC Pikfyvef/f, and age-matched wild-type (WT) mice. Box represents 25th and 75th percentiles; whiskers represent the range. (One-way ANOVA with Dunnett’s.). h. Percentage of pancreas occupied by normal tissue as determined by histological analyses of KC Pikfyve+/+, KC Pikfyvef/+, and KC Pikfyvef/f mice at 40 weeks of age. Box represents 25th and 75th percentiles; whiskers represent the range. (One-way ANOVA with Dunnett’s.). i. Breeding design for the generation of KPC Pikfyve+/+ and KPC Pikfyvef/f mice. Mouse cartoon was adapted from Adobe Stock Image (Asset #304271210). j. Representative images of PIKfyve BaseScope staining from pancreas tissue of 25-week-old KPC PIKfyve+/+ and KPC PIKfyvef/f mice. Scalebar = 20 μm. k. In situ Pikfyve levels in KPC Pikfyve+/+ and KPC Pikfyvef/f murine pancreas lesion vs normal tissue as determined by BaseScope. The KPC Pikfyve+/+ scores used as a reference are the same as those used in Fig. 1b. The two cohorts were stained and analyzed in the same batch. Box represents 25th and 75th percentiles; whiskers represent the range. (Multiple unpaired two-tailed t-test.). l. The age at death of mice in KPC PIKfyve+/+ and KPC PIKfyvef/f cohorts that were analyzed in Fig. 1f,g. Box represents 25th and 75th percentiles; whiskers represent the range. (Unpaired two-tailed t-test.). Statistics and reproducibility: b. n = 4 sections taken from individual KC Pikfyve+/+ animals analyzed for both normal and lesional areas; n = 5 sections taken from individual KC Pikfyvef/+ animals analyzed for both normal and lesional areas; n = 3 KC Pikfyvef/f sections taken from individual animals analyzed for normal and lesional areas. P-values: KC Pikfyve+/+ normal vs KC Pikfyvef/f normal:1.1e-5; KC Pikfyve+/+ lesion vs KC Pikfyvef/f lesion:3.1e-6. c. KC Pikfyve+/+ n = 4, KC Pikfyvef/+ n = 6, KC Pikfyvef/f n = 3, WT n = 3 individual animals. P-values: KC Pikfyve+/+ vs KC Pikfyvef/+:1.4e-8; KC Pikfyve+/+ vs KC Pikfyveff+:1.5e-8; KC Pikfyve+/+ vs WT:1.0e-8. d. KC Pikfyve+/+ n = 4, KC Pikfyvef/+ n = 6, KC Pikfyvef/f n = 3, WT n = 3 individual animals. P-values: KC Pikfyve+/+ vs KC Pikfyvef/+:2.1e-8; KC Pikfyve+/+ vs KC Pikfyvef/f:1.6e-8; KC Pikfyve+/+ vs WT:4.2e-9. e. KC Pikfyve+/+ n = 4, KC Pikfyvef/+ n = 6, KC Pikfyvef/f n = 3, WT n = 3 individual animals). f. These images are representative of n = 4 KC Pikfyve+/+; n = 6 KC Pikfyvef/+, n = 3 KC Pikfyvef/f; n = 3 WT. g. KC Pikfyve+/+ n = 7, KC Pikfyvef/+ n = 4, KC Pikfyvef/f n = 5, WT = 3 individual animals. h. KC Pikfyve+/+ n = 7, KC Pikfyvef/+ n = 4, KC Pikfyvef/f n = 5, WT = 3 individual animals. k. The KPC normal had n = 8 individual animals; the KPC lesion n = 14 individual animals; 8 animals were shared between these two groups. The KPC Pikfyvef/f normal group had n = 11 individual animals, and the KPC Pikfyvef/f lesion n = 16 individual animals; 11 animals were shared between these two groups. P-values: KPC Pikfyve+/+ normal vs KPC Pikfyve+/+ lesion:3.6e-5; KPC Pikfyve+/+ normal vs KPC Pikfyvef/f normal:1.9e-8; KPC Pikfyve+/+ lesion vs KPC Pikfyvef/f lesion: 1.6e-14. l. KPC PIKfyve+/+ n = 15, KPC PIKfyvef/f n = 16 individual animals.
