Fig 2: Raptor is not required for tuft cell differentiation in intestinal organoids. Vilin-Cre ER (-); Raptor (f/f) (WT, n = 3) and Vilin-Cre ER (+); Raptor (f/f) (i-Raptor-/-, n = 3) mice were injected intraperitoneally with TAM for 3 consecutive days, fed with liquid diet for 7 days and the small intestine removed at the end of the treatment. Crypts were isolated, mixed with matrigel and cultured into organoids. Organoids were then passaged and stimulated with 50 ng/ml IL-4 for 48 hours. (a) Representative whole-mount immunofluorescence shows tuft cells in green using DCLK1 antibody. (b) Western blot assay demonstrated DCLK1 expression after IL-4 stimulation. Raptor deletion in the organoid was also confirmed. GAPDH was used as loading control. (c) DCLK1 and IL-25 mRNA expression was measured via Real-time PCR assay.

Fig 3: DCLK1 silencing suppresses the viability, migration and invasion of nasopharyngeal carcinoma cells. si-DCLK1 or si-NC were transfected into 5-8F and 6-10B cells. (A) Effect of DCLK1 silencing on DCLK1 expression in 5-8F and 6-10B cells as determined via reverse transcription-quantitative PCR. (B) Proliferative activity of 5-8F and 6-10B cells as determined by Cell Counting Kit-8 assay. (C and D) Effect of DCLK1 silencing on the migration and invasion of 5-8F and 6-10B cells as determined using Transwell assays (magnification, ×200). *P<0.05 vs. si-NC. DCLK1, doublecortin-like kinase 1; si, small interfering RNA; NC, negative control.

Fig 4: The minimal KLHL15 docking region of DCLK2 includes both DCX domains.A, domain diagrams of DCLK2, DCLK3, and full-length or truncated DCLK chimeras. Red arrows mark the locations of the FRY sequence. B, GFP-tagged KLHL15 was coexpressed with the indicated FLAG-tagged DCLK constructs in COS-1 cells, and interactions were analyzed by GFP-IP and probing for FLAG. Shown are Western blots representative of four independent experiments. Molecular mass marker positions are shown in kilodaltons. FL, full length; Ub, polyubiquitinated species.

Fig 5: liquid diet is sufficient to eliminate Tm in mice. Vilin-Cre ER (-); Raptor (f/f) (WT, n = 4 per group) mice were subject to 50% proximal SBR or Sham operation, small intestinal tissue and stool was collected at the time of surgery and different days after surgery as indicated. (a) Representative IHC shows tuft cells in brown using anti-DCLK1 antibody. POD14 indicates postoperative day 14. (b) Real-time PCR quantification of Tm burden in collected stool as indicated.