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. 2019 Dec 6;2019:9186469. doi: 10.1155/2019/9186469

Figure 2.

Figure 2

Artificially induced and endogenous ALCAT1 levels. (a) Determination of relative ALCAT1 levels in HeLa cells supplied with different sugars and in cells transiently transfected with ALCAT1-EGFP and ALCAT1-HA-tag, respectively, 48 h after transfection. The transfection efficiency usually was ~60%; the data shown is not corrected for this. The upper band shows the tagged ALCAT1 version, the lower band the endogenous ALCAT1. VDAC, a mitochondrial protein, was used as loading control. For the quantification, ALCAT1 was normalized on the VDAC level. Total ALCAT1 and endogenous ALCAT levels were determined. A-GFP: ALCAT1-2A-GFP-NSL, A-HA: ALCAT1-HA-tag. N = 3 biological replicates. Left graph: total ALCAT levels, mean values are indicated. Right graph: only endogenous ALCAT levels are shown. Box and whisker plots. Statistics: ANOVA with post hoc Tukey. (b) Influence of metabolic conditions on endogenous ALCAT1 levels. The quantification includes values from (a). N = 8 technical replicates from n > 2 biological preparation for ALCAT in low glucose (LGlc; 5.6 mM), N = 8/n = 3 for galactose (Gal) 2 d; N = 4/n = 3 for galactose 4-5 d; N = /n = 2 for high glucose (HGlc, 25 mM). VDAC was used as a loading control. Statistics: SD are shown, ANOVA with post hoc Fisher LSD test. n.s.: no significant differences of the mean values.