Fig 2: IP3R-Grp75-VDAC1-MCU axis, mitochondrial Ca2+, and apoptosis in podocytes treated with ADR and Ang II. Ctl, control; ADR, Adriamycin; Ang II, angiotensin II; IP3R, inositol 1,4,5-triphosphate receptor; Grp75, glucose-regulated protein 75; VDAC1, voltage dependent anion channel 1; MCU, mitochondrial calcium uniporter; a, compared with the Ctl group; **, P < 0.01; *, P < 0.05. Compared with the Ctl podocytes, a) The cells in Q3 which were annexin high and PI low were counted as apoptotic cells. Significantly increased apoptosis rate was found in ADR- (n = 3) and Ang II-treated podocytes (n = 3), b) Significantly increased levels of active caspase-3 was found in podocytes treated with ADR (n = 4) and Ang II (n = 4), c) Mitochondrial Ca2+ levels were increased in podocytes treated with ADR (n = 12) and Ang II (n = 12), d) Significantly increased expression of IP3R, Grp75, VDAC1, and MCU were found in podocytes treated with ADR (n = 6) and Ang II (n = 6). e Co-IP were performed to analyze the IP3R-Grp75-VDAC1 interaction. Normal rabbit IgG without antigenicity was used as a negative control. Lysates from both Ctl and ADR- or Ang-II treated podocytes without immunoprecipitation were used as a positive control (input). The proteins pulled down by anti-Grp75 antibodies was analyzed by western blotting. Compared with the Ctl podocytes, there was a significant increase in the amount of IP3R, Grp75, and VDAC1 in pulled down samples from ADR- (n = 3) or Ang II-treated podocytes (n = 3)

Fig 3: ER stress homeostasis regulated by NB-UVB/ADSCs transplantation treatment. (A) Oxidative stress level detected by ELISA. (B) ROS detected by immunofluorescence (scale bar=20 µm). (C) The relative expression level of protein (GRP78 and caspase-12) detected by western blotting. (D) GRP75 expression level detected by western blotting. All the data are represented as the mean ± standard deviation (n=3). *P<0.05. ER, endoplasmic reticulum; NB-UVB, narrow band ultraviolet B; ADSCs, adipose-derived stem cells; ROS, reactive oxygen species; GRP, glucose-regulated protein; Con, mice were treated with Vaseline only; VIT, mice in vitiligo model group treated with 40% monobenzone cream; NUA, mice treated with narrow band ultraviolet B and adipose-derived stem cells together.

Fig 4: MKT-077 potentiates 17-AAG induced HCC cells apoptosis.(A) The indicated cell lines were treated with MKT-077 (MKT) or 17-AAG alone or in combination at indicated dosages for 24 hours. Cell viability was qualified using the Cell Counting Kit-8 (Dojindo Laboratories). * denotes P < 0.05 comparing 17-AAG (1 µM or 5 µM) to MKT-077 (3 µg/mL) +17-AAG (1 µM or 5 µM). (B) Increased cell apoptosis following combined treatment. Bel-7402, HuH7 and Hep3B cells treated as described above were subjected to Annexin-V and PI staining and cell apoptosis was quantified by FACS. * denotes P < 0.05 comparing 17-AAG (1 µM or 5 µM) to MKT-077 (3 µg/mL) +17-AAG (1 µM or 5 µM). (C) Levels of cleaved PARP, GRP75, Akt, phospho-AktSer473, phosphor-AktThr308, p53, phospho-p53Ser15, and phospho-p53Ser37 were detected by Western Blotting analysis. GAPDH served as a loading control.

Fig 5: Knocking down Grp75 protected podocytes from ADR- and Ang II-induced apoptosis in vitro. Ctl, control; Ang II, angiotensin II; Grp75, glucose-regulated protein 75; siCtl, non-targeted negative control siRNA; siGrp75, siRNA to knock down Grp75; a, compared with the Ctl group; b, compared with the ADR or Ang II group; *, P < 0.05; **, P < 0.01. a Knocking down Grp75 decreased Grp75 compared with non-targeted negative control siRNA group; b) The cells in Q3 which were annexin high and PI low were counted as apoptotic cells. Compared with ADR or Ang II -treated podocytes, knock down of Grp75 decreased the podocyte apoptosis rate significantly. c Compared with ADR or Ang II -treated podocytes, knock down of Grp75 decreased active caspase-3 significantly. d Compared with ADR or Ang II -treated podocytes, knock down of Grp75 decreased mitochondrial Ca2+ levels significantly