Skip to main content
. 2022 Apr 22;25(5):104282. doi: 10.1016/j.isci.2022.104282

Figure 1.

Figure 1

Her2 transformed cells showed upregulation of UPR genes in response to JG98

Transformation with Her2 oncogene enhances the sensitivity of the MCF10A cells to JG-98 treatment. Transcriptome analysis was performed with untransformed and transformed MCF10A cells treated with 1 μM JG-98 treatment for 12 h or left untreated.

(A) GSEA analysis of RNAseq data shows significant upregulation of UPR pathway following JG-98 treatment.

(B) Hallmark genes of unfolded protein response are significantly enriched in transformed cells in response to JG-98 treatment compared to untransformed cells. Color key represents the raw Z score. The experiments were performed in biological duplicates (n = 2).

(C) Sensitivity of transformed and untransformed cells to JG-98. MCF10A or Her2-transformed MCF10A cells were treated with 1 μM JG-98 for 24 h at 50% confluency or left untreated. Cell viability was counted by number of DAPI stained cells using Hermes Imaging systems (representative images shown). Experiment was performed in biological replicates (n = 3) and high throughput analysis was done using 31 images from each well using in-built Athena software package.

(D) Scatterplot showing the relative percentage of cells survival after 24 h of JG-98 treatment with means ± SEM (Quantification of data shown in (C)). Statistical analysis was performed using two-way ANOVA.

(E) Efficiency of CHOP (DDIT3) depletion following siRNA treatment. Levels of CHOP mRNA were quantified by qPCR, experiment was performed in biological replicates (n = 3), scatterplot showing the relative mRNA depletion represented as means ± SEM, statistics was performed using unpaired Welch’s correction, two-tailed t-test.

(F) CHOP depletion reduces cell death in response to JG-98. Cells were transfected with siControl or siCHOP, followed by 1μM of JG-98 treatment for 24 h or left untreated. Cell viability was counted by number of DAPI stained cells using Hermes Imaging systems and representative images shown, experiment was performed in biological replicates (n = 3) and analysis was done using 126 images from each well.

(G) Scatterplot showing relative percentage of cells survival after 24 h of JG-98 treatment plotted as means ± SEM (Quantification of data shown in (F)), Statistical analysis was performed using two-way ANOVA. All statistical analysis was performed using Graphpad (v9), level of significance was taken as (∗p < 0.0332, ∗∗p < 0.0021, ∗∗∗p < 0.0002). See also Figures S1S2, and Tables S1–S3.