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. Author manuscript; available in PMC: 2021 Sep 3.
Published in final edited form as: Biochem Biophys Res Commun. 2020 Jul 30;529(4):1011–1017. doi: 10.1016/j.bbrc.2020.07.002

Figure 2. LY83583 stimulated Type I collagen secretion depends on P4HA1 and P4HA2 activity but not their expression level.

Figure 2.

(a) Addition of LY83583 did not result in significant differences in P4ha1 or P4ha2 mRNA level within 6 hrs based on RT-qPCR. (b) Immunoblotting with P4HA1 or P4HA2 specific antibody showed that LY83583 did not enhance the levels of these two enzymes. In fact, significantly less P4HA1 or P4HA2 was detected by immunoblotting 6 hrs after LY83583 addition. P4HA1/actin or P4HA2/actin was quantified by IMAGE J software by arbitrary defining the signal at 0 hr after addition of LY83583 as 1. (c, d and e) Silencing P4ha1 (c) or P4ha2 (d) greatly decreased the amount of pepsin-resistant Type I collagen in the culture medium (panel 1) but slightly enhanced the level of P4HA2 (c-panel 3) or P4HA1 (d-panel 4). Silencing both p4ha1 and p4ha2 (e) totally blocked pepsin-resistant Type I collagen in the culture medium. Pepsin-resistant collagen in the culture medium to actin was quantified by IMAGE J software and is depicted graphically. * indicated pepsin-resistant col 1 α2. N.s.: not statistically significant. P values were indicated in each panel if available.