Fig. 2: hIAPP/sIAPP oligomer clearance by MSL-7. | Nature Communications

Fig. 2: hIAPP/sIAPP oligomer clearance by MSL-7.

From: An autophagy enhancer ameliorates diabetes of human IAPP-transgenic mice through clearance of amyloidogenic oligomer

Fig. 2: hIAPP/sIAPP oligomer clearance by MSL-7.The alternative text for this image may have been generated using AI.

a Prepro-mIAPP-HA-transfected or Prepro-hIAPP-HA-transfected INS-1 cells were treated with MSL-7 in the presence or absence of bafilomycin A1 (BAF). Lysate was subjected to immunoblotting using anti-HA or anti-β-actin antibody (ACTB). Numbers indicate fold changes of hIAPP dimer normalized to ACTB bands. (Con, control-transfected). b Prepro-mIAPP-HA-transfected or Prepro-hIAPP-HA-transfected cells were treated with MSL-7 in the presence or absence of 3-MA. Oligonucleosome content was determined (F = 19.5, df treatment = 11, df residual = 36). c–d MSL-7-induced nuclear translocation of TFEB (c) and TFE3 (d) in monkey islet cells was counted (t = 33.2, df = 14 in c; t = 45.7, df = 14 in d) (right of c, d). Representative pictures are presented (left of c, d). e mRFP-GFP-LC3-transfected monkey islet cells were treated with MSL-7. The numbers of autophagosome (t = 9.5, df = 44) and autophagolysosome (t = 10.9, df = 44) were counted (right). Representative pictures are presented (left). Inset images were magnified to show red (autophagolysosomes) and yellow (autophagosomes) puncta. f Monkey islet cells treated with MSL-7 in the presence or absence of 3-MA were subjected to immunostaining using A11 antibody. The number of A11+ puncta was counted (F = 86.3, df treatment = 2, df residual = 42) (right). Representative pictures are presented (left). g Monkey islet cells treated with MSL-7 in the presence of BAF, cells were immunostained using A11 and anti-LC3 antibodies. The number of LC3 puncta colocalized with A11+ oligomer was counted (t = 13.9, df = 38) (right). Representative pictures are shown (left). Inset images were magnified to shows LC3 puncta colocalized with A11+ oligomer. h Apoptosis was determined in MSL-7-treated monkey islet cells in the presence or absence of 3-MA (F = 10.1, df treatment = 3, df residual = 20). All data in this figure are the means ± SEM from more than 3 independent experiments performed in triplicate. (scale bar, 5 μm) *P < 0.05; **P < 0.01; ***P < 0.001 by one-way ANOVA with Tukey’s test (b, f, h) and two-tailed Student’s t-test (c, d, e, g). Source data are provided as a Source Data file.

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