Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2024 Oct 29;221(11):e20231832. doi: 10.1084/jem.20231832

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2024 Hsu et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).

PMC Copyright notice

Figure 8. — IMPA1/inositol/IMPDH2 maintains PCSCs with AR^low/− features. (A and B) Immunoblotting of ALDH^low/− and ALDH^high cells sorted from LNCaP (A) and PC3 (B) cells stably expressing shLuc or shIMPA1 (#1 and #2) with indicated antibodies. Immunoblotting data were verified in at least two independent experiments. (C and D) Relative gene expression of reprogramming factors and epigenetic regulators in 22RV1 cells stably expressing shLuc, shIMPA1, or shIMPDH2 were determined by qRT-PCR analysis. Data are shown as the mean ± SEM of three independent experiments for each group. **, P < 0.01; ***, P < 0.001 by two-tailed unpaired t test. (E) Relative gene expression of AR and AR target genes in 22RV1 cells stably expressing shLuc or shIMPA1 upon 40 μM of inositol (Ins) treatment for 48 h were determined by qRT-PCR. Data are shown as the mean ± SEM of three independent experiments for each group. *, P < 0.05; **, P < 0.01; ***, P < 0.001 by two-tailed unpaired t test. (F) Relative gene expression of AR and AR target genes in 22RV1 cells stably expressing shLuc or shIMPDH2 (#1 and #2) were determined by qRT-PCR. Data are shown as the mean ± SEM of three independent experiments for each group. *, P < 0.05; **, P < 0.01 by two-tailed unpaired t test. (G) Relative gene expression of SOX2, Nanog, Oct4 and SYP in 22RV1 cells upon vehicle or 500 μM of LiCl treatment for 48 h were determined by qRT-PCR. Data are shown as the mean ± SEM of three independent experiments for each group. *, P < 0.05; **, P < 0.01; ***, P < 0.001 by two-tailed unpaired t test. (H) Immunoblotting of 22RV1 cells stably expressing shLuc or shIMPA1 (#1 and #2) with indicated antibodies. Immunoblotting data were verified in at least two independent experiments. (I) Immunoblotting of 22RV1 cells stably expressing shLuc or shIMPDH2 (#1 and #2) with indicated antibodies. Immunoblotting data were verified in at least two independent experiments. (J) Immunoblotting of tumor organoid from TRAMP mice expressing shLuc or shIMPA1 (#1, #2, and #3) with indicated antibodies. Immunoblotting data were verified in at least two independent experiments. (K) Immunoblotting of 22RV1 cells expressing shLuc or shIMPA1 upon 25 μM of inositol (Ins) treatment with indicated antibodies. Immunoblotting data were verified in at least two independent experiments. (L) IHC of 22RV1 xenograft tumors stably expressing shLuc or shIMPA1 with indicated antibodies. IHC data were verified in at least two independent experiments. Scale bar, 100 μm. (M) IHC of prostate tissue from TRAMP/PB-Cre4 and TRAMP/Impa1^FL/FL/PB-Cre4 mice at the age of 7 mo with indicated antibodies. IHC data were verified in at least two independent experiments. Scale bar, 50 μm. (N and O) Relative gene expression of SOX2, Nanog, Oct4, and AR genes in 22RV1 cells upon the treatment of 40 μM of inositol (Ins), 40 μM of guanosine (Gua), or 40 μM of adenosine (Ade) for 48 h were determined by qRT-PCR. The data are shown as the mean ± SEM of three independent experiments for each group. *, P < 0.05; **, P < 0.01; ***, P < 0.001; NS, non-significant by two-tailed unpaired t test. (P) Relative gene expression of luminal markers in LNCaP cells upon the treatment of 40 μM of inositol (Ins), 40 μM of guanosine (Gua), or 40 μM of adenosine (Ade) for 48 h were determined by qRT-PCR. The data were shown as the mean ± SEM of three independent experiments for each group. *, P < 0.05; **, P < 0.01; NS, non-significant by two-tailed unpaired t test. (Q) Relative gene expression of lineage markers in 22RV1 cells upon the treatment of vehicle (Veh), 40 μM of inositol (Ins), 40 μM of guanosine (Gua), or 40 μM of adenosine (Ade) for 48 h, followed by qRT-PCR. (R) Relative gene expression of lineage markers in 22RV1 cells stably expressing shLuc or shIMPA1 upon the treatment of vehicle (Veh), 40 μM of inositol (Ins), or 40 μM of guanosine (Gua) for 48 h, followed by qRT-PCR. (S) Relative gene expression of lineage markers in 22RV1 cells stably expressing shLuc or shIMPDH2 upon the treatment of vehicle (Veh), 40 μM of inositol (Ins), or 40 μM of guanosine (Gua) for 48 h, followed by qRT-PCR. All data in Q–S are shown as the mean ± SEM of three independent experiments for each group. *, P < 0.05; **, P < 0.01; ***, P < 0.001, NS, non-significant by two-tailed unpaired t test. Source data are available for this figure: SourceData F8.