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Fig. 3 | Respiratory Research

Fig. 3

From: Dysfunctional lactate metabolism in human alveolar type II cells from idiopathic pulmonary fibrosis lung explant tissue

Fig. 3

Relationship of LDH isoenzyme expression to oxidative and glycolytic metabolism in human IPF AEC2s. Purified AEC2s from tissues sections of three IPF patients were subjected to targeted siRNA transfection to knockdown LDHA expression (LDHA KD; compared to cells transfected with a scrambled siRNA control). A LDHA mRNA expression in IPF AEC2 cells 72 h after transfection was measured using qPCR. Fold-changes of LDHA normalized to 18S rRNA were calculated using ΔΔCt analysis and values for triple replicates were averaged; relative LDHA expression in scrambled siRNA control cells was set as 1.0. B Non-denatured lysates from IPF AEC2 72 h after transfection were separated by native gel electrophoresis and subjected to colorimetric staining for LDH activity to measure the % of each isoenzyme tetramer (LDH1 to 5) comprising total LDH. C OCR rates were measured and compared between these transfected IPF AEC2s cultured in the presence of both glucose and lactate. D The ratio of PPR to OCR in these transfected cells. All values were normalized to total cellular protein

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