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. Author manuscript; available in PMC: 2021 Jan 1.
Published in final edited form as: FASEB J. 2019 Nov 28;34(1):930–944. doi: 10.1096/fj.201902155R

Figure 7. Pharmacological inhibition and molecular knockdown of PKM2 attenuate high glucose-induced transglutaminase 2 activity and fibrogenesis.

Figure 7.

Pulmonary artery adventitial fibroblasts were starved overnight in glucose-free media followed by stimulation with normal (NG; 5 mM) or high (HG; 50mM) glucose media for 72 hours. (A) Fibroblasts were exposed to NG or HG media after pretreatment with vehicle control or Shikonin (Shi; 0.5 – 5 μM). Representative Western blot images of protein extracts showing serotonylated fibronectin (sFn; 220 kDa), transglutaminase 2 (TG2; 78 kDa), α-smooth muscle actin (α-SMA; 42 kDa) and β-actin (42 kDa) expression. Bar graph demonstrating fold change in expression normalized to β-actin by densitometry analysis. n=4–6 replicates/ group; mean±SEM. Statistical analysis was performed by One Way ANOVA Tukey post-hoc test. p<0.05 *compared to NG+vehicle; #compared to HG+vehicle. (B) Fibroblasts were exposed to NG or HG media for 72-hours after transfection with control siRNA or pyruvate kinase muscle isoform 2 (PKM2) siRNA as described in methods section. Representative Western blot images of protein extracts showing PKM2 (58 kDa), sFn (220 kDa), type 1 collagen (Col1; 145 kDa), TG2 (78 kDa) and β-actin (42 kDa) expression. Bar graph demonstrating fold change in expression normalized to β-actin by densitometry analysis. n=4 replicates/ group; mean±SEM. Statistical analysis was performed by One Way ANOVA Tukey post-hoc test. p<0.05 *compared to NG+control siRNA; #compared to HG+control siRNA.