Additional file 2 of LASS2 enhances chemosensitivity to cisplatin by inhibiting PP2A-mediated β-catenin dephosphorylation in a subset of stem-like bladder cancer cells
Creators: name: Shi, Hongjin
name: Tan, Zhiyong
name: Duan, Bowen
name: Guo, Chunming
name: Li, Chong
name: Luan, Ting
name: Li, Ning
name: Huang, Yinglong
name: Chen, Shi
name: Gao, Jixian
name: Feng, Wei
name: Xu, Haole
name: Wang, Jiansong
name: Fu, Shi
name: Wang, Haifeng
Publisher: name: figshare
Year: 2024
Subjects: subject: Biochemistry
subject: Medicine
subject: Cell Biology
subject: Genetics
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Pharmacology
subject: Chemical Sciences not elsewhere classified
subject: Cancer

description: Additional file 2: Fig. S1. High-resolution images of immunohistochemical staining. Fig. S2. A, Kaplan–Meier curve and Log-rank test of overall survival for patients received cisplatin-based chemotherapy in GSE70691 and GSE69795 datasets. B, The expression levels of LASS2 and stemness-related genes (CD44, STAT3, CD47, ALDH1A1) in different molecular subtypes of MIBC. C and D, The expression levels of LASS2 in different molecular subtypes of breast cancer. *p &lt; 0.05, determined by the Kruskal-Wallis test. Fig. S3. A, Left panel: Heatmap showing the correlation coefficient of each bladder cancer cell line for basal-like (BL) or non-basal-like (nBL) bladder cancer in GSE64572. Right panel: mRNA expression levels of CD44 in bladder cancer cell lines. B, qPCR analysis of CD44 expression in the indicated cell lines. C, Cisplatin IC50 vales of the indicated cell lines. D, Scatter plot showing the correlation between LASS2 expression levels and cisplatin IC50 values. r, Spearman correlation coefficient. E, The overexpression and knockdown efficiency of LASS2 was verified by western blot. F, Flow cytometry analysis of the percentage of CD44+ALDH1A1+ subpopulation in bladder cancer cell lines. G, Quantitative analysis of ceramides by LC–MS in chemoresistant and chemosensitive bladder cancer specimens. H, Pathway activity score of LASS2-high group versus LASS2-low group is presented though bubble color and size. The bubble color from green to red represents the pathway activity from inhibition to activation, and the bubble size is positively correlated with the FDR value. Fig. S4. A, Cell viability and cisplatin IC50 assay of cell lines from different cancer types. B, Table showing the cisplatin IC50 values in cell lines from different cancer types. C, Cell viability and IC50 assay of the 5637 cell line treated with different chemotherapy agents. Fig. S5. BCSC sorting and examination. A, The gating strategies of FACS sorting. B, Percentages of CD44+ALDH1A1−, CD44−ALDH1A1+, CD44+ALDH1A1+, and CD44−ALDH1A1− subpopulations in BCSCs and BCNSCs. C, In vitro extreme limiting dilution assay of BCSCs and BCNSCs. D, qPCR analysis of CSC markers in BCSCs and BCNSCs. E-H, The indicated numbers (2×101, 2×102, 2×103 and 2×104) of BCSCs and BCNSCs were subcutaneously injected into nude mice. Tumor images (E), tumor incidence (F), and tumor weight (G, H) are shown. I, Quantitative analysis of ceramides by LC–MS in BCSCs and BCNSCs. J, Differentially expressed resistance-related genes in BCSCs were identified by an RT2 Profiler PCR Array (p &lt; 0.05 and fold change &gt; 5 genes were selected). Data are presented as the mean ± SD of at least three technical replicates. *p&lt;0.05, **p&lt;0.01, ***p&lt;0.001, determined by Student’s t test. Fig. S6. A, Cisplatin sensitivity analysis of PDX tumors. The IC50 values of cisplatin for each tumor are presented in the plot. B, Percentage of CD44+ALDH1A1+ subpopulation in PDX tumors. C. Immunohistochemistry staining of CD44 in the indicated PDX tumors. Fig. S7. A, Measurement of mouse body weight during treatment. B, Hematological parameters of mice in different groups. C, Organ weight of mice in different groups. Fig. S8. Analysis of the CERS family genes. A, Heatmap showing the mRNA expression of CERS family genes in each cancer. B, qPCR analysis of CERS family genes in chemosensitive and chemoresistant bladder tumors. C, Pie plot showing the constitute of heterozygous/homozygous copy number variation (CNV) of CERS family genes in each cancer. D, Correlation analysis of CNV with mRNA. The mRNA expression data and CNV raw data were merged by TCGA barcode, followed by Spearman correlation analysis. Blue bubbles represent negative correlations, with red bubbles representing positive correlations. The bubble size is positively correlated with the FDR significance. The black outline border indicates FDR ≤ 0.05. Fig. S9. Original blot images.