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

. 2019 Oct 7;9:14394. doi: 10.1038/s41598-019-50840-7

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

© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Effects of HSPA1 or HSPA2 depletion on the proliferation, clone-forming ability and chemoresistance of NCI-H1299 cells. (A,B) Levels of HSPA proteins in wild-type (wt); control sh-luc cells stably transduced with a non-targeting shRNA-luc sequence; sh-A1.N and sh-A1.S cell lines stably transduced with HSPA1-targeting sh-RNA-A1.N or sh-RNA-A1.S sequences, respectively (A); sh-A2.3 and sh-A2.4 cell lines stably transduced with HSPA2-targeting sh-RNA-A2.3 or sh-RNA-A2.4 sequences, respectively (B). Representative immunoblots are shown (n = 3), actin was used as a protein loading control. Graph shows results of densitometric analysis of HSPA1 (A) or HSPA2 (B) immunodetection (HSPA1 n = 3, HSPA2 n = 7). (C) Cell cycle phase distribution in sub-confluent cells at 48 hours (h) after plating. Graph shows percentage of cells (mean ± SD, n = 3, each in two technical replicas). (D) Cell proliferation assessed by MTS assay. Results are expressed as mean ± SD (n = 3, each in three technical replicas) in relation to values obtained at 24 h after plating. (E) Results of proliferation assay (n = 3, each in six technical replicas) assessed using crystal violet staining. Relative absorbance of stained cells was plotted against time (24–120 h) of continuous growth. (F) Number of colonies formed by cells plated onto 6-well dishes (1 × 10³ cells/well) and cultured for 7–8 days. Colonies were counted manually (mean ± SD, n = 5 each in three technical replicas). (G,K) Effects of HSPA1 or HSPA2 depletion on resistance of cells to cisplatin (CDDP) (G,H), carboplatin (CPT) (I) or bortezomib (BTZ) (J-K). Cell viability was measured using MTS assay after 72 h treatment (G,I,J). Results are expressed relative to untreated control (mean ± SD from at least three independent experiments, each in triplicate, *p < 0.05, statistical significance was determined by two-tailed t-test). Cell death detection using propidium iodide (PI) uptake test after 24 h treatment with CDDP (H) or BTZ (K) and/or following 24 or 48 h growth without drugs. Results show mean values ± SD from two (H) or three (K) independent repeats, each at least in duplicate. Statistical significance was determined using two-tailed t-test.