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

. 2021 May 6;129(5):057003. doi: 10.1289/EHP7612

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

EHP is an open-access journal published with support from the National Institute of Environmental Health Sciences, National Institutes of Health. All content is public domain unless otherwise noted.

PMC Copyright notice

Figures 3A, 3C, 3D, and 3E are western blots plotting 0, 0.05, 0.1, 0.2, 0.4, and 0.8 (milligram per milliliter) (columns) across inositol-requiring transmembrane kinase or endoribonuclease 1 lowercase alpha, Activating transcription factor 6, phosphorylation of eukaryotic initiation factor-2 lowercase alpha, eukaryotic initiation factor-2 lowercase alpha, and glyceraldehyde-3-phosphate dehydrogenase; Cytosolic phospholipase A 2, Cyclooxygenase 1, and glyceraldehyde-3-phosphate dehydrogenase; Phosphorylated-mammalian target of rapamycin, mammalian target of rapamycin, Phosphorylated-Protein kinase B, Protein kinase B, and glyceraldehyde-3-phosphate dehydrogenase; and Sequestosome-1, Beclin-1, light chain 3-1, light chain 3-2, and glyceraldehyde-3-phosphate dehydrogenase (rows), respectively. Figure 3B is a western blot plotting 0, 5, 10, 20, 30, and 60 minutes (columns) across Phosphorylated-Extracellular signal-regulated kinases 1 and 2, Extracellular signal-regulated kinases 1 and 2, Phosphorylated-c-Jun N-terminal kinase, c-Jun N-terminal kinase, Phosphorylated-p 38 mitogen-activated protein kinases, p 38 mitogen-activated protein kinases, and glyceraldehyde-3-phosphate dehydrogenase (rows). Figure 3F is a display of six columns, namely, 0, 0.05, 0.1, 0.2, 0.4, and 0.8 (milligram per milliliter) and one row, namely, Polystyrene microplastics. Figure 3G is a bar graph plotting light chain 3-1 positive cells percentage, ranging from 0 to 40 in increments of 10 (y-axis) across Polystyrene microplastics (0, 0.05, 0.1, 0.2, 0.4, and 0.8 milligram per milliliter) (x-axis). — ER stress-related proteins, MAPK signaling pathways, inflammation-related proteins, AKT/mTOR signaling pathways, and autophagosome-related proteins in HK-2 cells treated with PS-MPs. (A) Representative western blot showing the expression of ER stress-related proteins $IRE 1 α$ , ATF6, $p -EIF 2 α$ , and $EIF 2 α$ after PS-MPs treatment at concentrations of 0.05, 0.1, 0.2, 0.4, and $0.8 mg / mL$ for 24 h. (B) Representative western blot showing the expression of MAPK signaling pathway components p-ERK1/2, ERK1/2, p-JNK, JNK, p-p38, and p38 after treatment with PS-MPs at $0.8 mg / mL$ for 0, 5, 10, 20, 30, and 60 min. (C) Representative western blot showing the expression of inflammation-related proteins ${cPLA}_{2}$ and COX-1 after PS-MP treatment at concentrations of 0.05, 0.1, 0.2, 0.4, and $0.8 mg / mL$ for 24 h. (D) Representative western blot showing the expression of AKT/mTOR signaling pathway components, such as p-mTOR, mTOR, p-AKT, and AKT after treatment with PS-MPs at concentrations of 0.05, 0.1, 0.2, 0.4, and $0.8 mg / mL$ for 1 h. (E) Representative western blot showing the expression of autophagy-related proteins p62, Beclin 1, and LC3, after PS-MP treatment at concentrations of 0.05, 0.1, 0.2, 0.4, and $0.8 mg / mL$ for 24 h. GAPDH served as an internal control. (F) LC3 expression (green) was detected after treatment with PS-MPs at concentrations of 0.05, 0.1, 0.2, 0.4, and $0.8 mg / mL$ for 24 h in immunostaining. DAPI (blue) was used for nuclear staining. $Scale bar: 25 μ m$ . (G) The results for LC3-positive cells were graphed and statistically analyzed. $n = 3$ . * $p < 0.05$ and *** $p < 0.001$ compared with the control group, as determined by one-way ANOVA, with Dunnett’s multiple comparison test. Significance: $0 -mg / mL$ group vs. $0.1 -mg / mL$ group: $p = 0.0265$ , $0 -mg / mL$ group vs. $0.2 -mg / mL$ group: $p < 0.001$ , $0 -mg / mL$ group vs. $0.4 -mg / mL$ group: $p < 0.001$ , $0 -mg / mL$ group vs. $0.8 -mg / mL$ group: $p < 0.0015$ . The data are presented as the $means \pm SDs$ . The western blotting results were quantified and statistically analyzed, as shown in Figures S3–S7. The mean and SD summary data for quantification of western blots are shown in Table S3. Note: AKT, protein kinase B; ANOVA, analysis of variance; ATF6, activating transcription factor 6; COX-1, cyclooxygenase-1; ${cPLA}_{2}$ , cytoplasmic phospholipase A2; DAPI, 4,6-dimidyl-2-phenylindole; $EIF 2 α$ , eukaryotic initiation factor 2 alpha; ERK1/2, extracellular signal-regulated kinases 1 and 2; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; HK-2 cells, human kidney 2 cells; $IRE 1 α$ , inositol-requiring enzyme $1 α$ ; JNK, c-Jun N-terminal kinase; MAPK, mitogen-activated protein kinase; mTOR, mitogen-activated protein kinase; p, phosphorylated; p-p38, phosphorylated-p38 mitogen-activated protein kinases; PS-MPs, polystyrene microplastics; SD, standard deviation.