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. Author manuscript; available in PMC: 2020 Jun 11.
Published in final edited form as: Nature. 2019 Dec 11;576(7787):477–481. doi: 10.1038/s41586-019-1831-x

Figure 4 |. SLC4A7 is necessary for Ras-induced macropinocytosis and tumor growth.

Figure 4 |

a, Quantification of TMR-dextran uptake following treatment of mutant Ras cells as shown. b, mRNA transcript levels of SLC4 family members in PDAC compared to normal adjacent (Adj) tissue from 74 patients (n=50 PDAC, n=24 adjacent normal). c, Effect of oncogenic Ras expression on SLC4A7 protein levels. Immunoblots of SLC4A7 in the plasma membrane (PM) fraction and whole cell lysate (WCL) from HeLa Ctl, HV12, or KV12 cells. d,e, Effect of doxycycline-inducible FLAG-KRasV12 (+Dox) expression in BxPC-3 cells on SLC4A7 expression and macropinocytosis. d, Immunoblot of SLC4A7 expression (vinculin, loading control) and (e) quantification of FITC-dextran uptake with or without Dox treatment following SLC4A7 knockdown in BxPC-3 cells. f, Immunoblot of SLC4A7 expression (vinculin, loading control) following KRas knockdown in MIA-PaCa-2 cells. g,h, Quantification of TMR-dextran uptake following SLC4A7 knockdown in (g) HeLa HV12 and (h) MIA-PaCa-2 cells treated as shown. i-j, Effect of dox-inducible SLC4A7 depletion in MIA-PaCa2 cells on macropinocytosis and tumor growth. i, Quantification of FITC-dextran uptake treated as shown. j, Waterfall plots of xenografts treated as shown relative to baseline. Each bar represents a tumor. Immunoblots (c,d,f) are representative of three biological replicates. ≥500 (a,e,g,h,i) cells were quantified in each biological replicate (n=3) with error bars indicating mean ±s.e.m.; p value: unpaired, two-tailed Student t-test. For c,d,f, gel source data located in Supplementary Figure 1.