Fig. 2
AQP9 expression, linked to CRC cell function, is low in KRASG12V CRC tissues and cells. A&B: Expression of AQP9 in KRASWT, KRASOTHER, and KRASG12V CRC tissues (Immunohistochemistry). (A) Microscopic results, (B) Quantitative results. (C) Expression of AQP9 in KRASWT, KRASOTHER, and KRASG12V CRC tissues (Western blot). (D) Construction of KRASG12V colorectal cancer cell lines using CRISPR/Cas9 RNP technology. (E) Sanger sequencing validation of KRASG12V CRC cell line. (F) Relative mRNA quantification of AQP9 in KRASWT and KRASG12V CRC cell. (G, H) Relative protein quantification of AQP9 in KRASWT and KRASG12V CRC cell. (G) Western blot, (H) Quantitative results. (I) Proliferation levels of KRASWT and KRASG12V CRC cell (CCK8 assay). (J) Invasion levels of KRASWT and KRASG12V CRC cell (Transwell assay). (K) Migration levels of KRASWT and KRASG12V CRC cell (Scratch assay). (L) Apoptosis levels of KRASWT and KRASG12V CRC cell (FITC-Annexin V/PI flow cytometry assay). (M) Alterations in glycerol concentration in the two cell types at various time points following the administration of exogenous glycerol. (B) n = 6, variance analysis; (F, H-M) n = 3, two independent sample t-tests.
