Fig. 4.

Downregulation of TMEM59L ameliorates N2a cell apoptosis induced by hydrogen peroxide insult. a, b, d Tmem59l-shRNA1, Tmem59l-shRNA2, or scrambled shRNA (sc-shRNA) were transfected into N2a cells for 48 h. a Cells were subjected to total RNA extraction, reverse transcribed, and qRT-PCR. Tmem59l mRNA levels were normalized to those of β-actin for comparison. Cells without transfection were used as a mock control. n = 3, ****p < 0.0001. b Equal amounts of cell lysates were subjected to Western blot analysis for LC3B-I/II. d LC3B-II and LC3B-I protein levels were quantified by densitometry for comparison. n = 3, *p < 0.05, **p < 0.01. c, e–h N2a cells were transfected with Tmem59l-shRNA1, Tmem59l-shRNA2, or sc-shRNA for 43 h and then treated with 300 µM H₂O₂ or not (control) for another 5 h. c Equal amounts of cell lysates were subjected to Western blot analysis for c-Caspase-3 and c-PARP. e c-Caspase-3 and f c-PARP levels were quantified by densitometry for comparison. n = 3, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. g Cells were stained by using TUNEL method and counterstained with DAPI. Scale bars, 100 µm. h TUNEL-positive cells were counted as apoptotic cells for comparison. ns, not significant, *p < 0.05, **p < 0.01