Extended Data Fig. 4: ZDHHC8 deficiency inhibits colorectal cancer growth in vivo.
a, Analysis of mRNA expression of ZDHHC8 in colon adenocarcinoma (COAD) and rectum adenocarcinoma (READ) using the Cancer Genome Atlas (TCGA) datasets. For COAD, n = 41 samples in normal tissues, and n = 458 samples in tumor tissues. For READ, n = 10 samples in normal tissues, and n = 166 samples in tumor tissues. b, ZDHHC8 mRNA expression level is negatively co-related with the infiltration of CD8+ T cells in COAD and READ based on TCGA database. c,d, mRNA and protein level of ZDHHC8 were detected by real-time PCR (c) or immunoblot analysis (d) in CRC patients. e, Immunoblotting analysis of GPX4 palmitoylation by ABE assay in CRC samples from patients. Adj, adjacent normal tissues; Ca, cancerous tissue. f, The cell viability of control (shCtrl) or Zdhhc8-knockdown (shZdhhc8) MC38 cells. g-i, Tumor images (g), tumor growth curve (h), and tumor weight (i) of shCtrl and shZdhhc8 MC38 subcutaneous tumors. j, Lipid peroxidation assessment of shCtrl and shZdhhc8 MC38 subcutaneous tumors. k, Flow cytometric analysis of CD8+ cells in shCtrl and shZdhhc8 MC38 subcutaneous tumors. l, Representative CD8α immunofluorescence staining images of MC38 tumor sections from shCtrl and shZdhhc8 groups (n = 6 mice per group, 2 sections per mouse). Scale bar, 50 μm. m-o, Flow cytometric analysis of IFNγ+ (m), TNF+ (n) and GZMB+ (o) CD8+ cytotoxic T cells in shCtrl and shZdhhc8 MC38 subcutaneous tumors. For a, the box represents the interquartile range (IQR). The central line indicates the median. The lower and upper bounds of the box represent the first quartile and the third quartile. The whiskers extend to the minima and maxima. Data points outside the range of the whiskers are considered outliers. For b, The Tumor Immune Estimation Resource (TIMER, https://cistrome.shinyapps.io/timer/) accomplished the immune infiltration analysis, and the tumor purity adjusted the results. Spearman’s rank correlation test is used, following a two-sided test without adjustment for multiple comparisons. The partial correlation coefficient (part.cor) and the corresponding p value are shown on the plot. n = 458 tumor samples for COAD, n = 166 tumor samples for READ. For c, data are presented as mean ± s.d. of n = 30 samples in normal group and n = 42 samples in tumor group. For d, quantification of the indicated protein level is determined by Image Lab software; data are presented as mean ± s.d. n = 4 independent samples per group. For e, quantification of the indicated protein level is determined by Image Lab software; data are presented as mean ± s.d. n = 3 independent experiments per group. For f, data are presented as mean ± s.d. of n = 4 independent samples per group. For h-k,m-o, data are presented as mean ± s.d. of n = 6 mice in each group. Statistical analysis was performed using two-tailed paired Student’s t-test (e), two-tailed unpaired Student’s t-test (a,c,d), one-way ANOVA with Dunnett’s test (i-k,m-o) and two-way ANOVA with Tukey’s test (h).
