Additional file 1 of Overexpression of HSF2 binding protein suppresses endoplasmic reticulum stress via regulating subcellular localization of CDC73 in hepatocytes
Creators: name: Zhang, Jia
name: Wang, Tao
name: Bi, Jianbin
name: Ke, Mengyun
name: Ren, Yifan
name: Wang, Mengzhou
name: Du, Zhaoqing
name: Liu, Wuming
name: Hu, Liangshuo
name: Zhang, Xiaogang
name: Liu, Xuemin
name: Wang, Bo
name: Wu, Zheng
name: Lv, Yi
name: Meng, Lingzhong
name: Wu, Rongqian
Publisher: name: figshare
Year: 2024
Subjects: Biochemistry
Medicine
Cell Biology
Molecular Biology
Chemical Sciences not elsewhere classified
Biological Sciences not elsewhere classified

description: Additional file 1: Table S1. The information of antibodies forwestern blot analysis. Table S2. The information of primers for q-PCR. Figure S1. HSF2BP expression is increased after tunicamycin-induced ER stress inmice. Liver H&amp;E staining (A) andits histological score (B) intunicamycin-induced ER stress. Original magnification, x100 and x200. C, Western blot analysis of HSF2BP and GRP78 in tunicamycin-induced ERstress. Quantitative analysis of GRP78 (D)and HSF2BP (E) intunicamycin-induced ER stress. Results are expressed as mean ± SE (n =4-6/group) and compared by one-way ANOVA. * p &lt; 0.05. Figure S2. HSF2BP expression is increased after tunicamycin-induced ERstress in cultured hepatocytes. A, Western blot analysisof HSF2BP and GRP78 in HL-7702 cells. Quantitative analysis of GRP78 (B) and HSF2BP (C) in HL-7702 cells. Results are expressed as mean ± SE (n =3/group) and compared by one-way ANOVA. * p &lt; 0.05. Figure S3. Theestablishment of hepatocyte-specific HSF2BP transgenic mice. A, The HSF2BP mRNA levels in variousorgans of HSF2BP transgenic (TG) mice. Western blotanalysis of HSF2BP (B) and itsquantitative analysis (C) in liverof hepatocyte-specific HSF2BP-TG and NTG mice. The immunohistochemical stainingof HSF2BP (D) and its IHC score (E) in liver of HSF2BP-TG and NTG mice.Original magnification, x400. F, Bodyweight of HSF2BP-TG and NTG mice. G, TheH&amp;E staining in liver of HSF2BP-TG and NTG mice. Original magnification, x400.H, The levels of HSFs (heat shock factors)and HSPs (heat shock proteins) in liver of HSF2BP-TG and NTG mice. I, The levels of inflammatory factorsin liver of HSF2BP-TG and NTG mice. Results are expressed as mean ± SE (n =4-6/group) and compared by t-test or one-way ANOVA. * p &lt; 0.05. Figure S4. Theestablishment of hepatocyte-specific HSF2BP knockout mice. A, The HSF2BP mRNA levels invarious organs of HSF2BP knockout (KO) mice. Western blot analysis of HSF2BP (B) and its quantitative analysis (C) in liver of HSF2BP-KO and WT mice.The immunohistochemical staining of HSF2BP (D)and its IHC score (E) in liver ofHSF2BP-KO and WT mice. Original magnification, x400. F, Body weight of HSF2BP-KO and WT mice. G, The H&amp;E staining in liver of HSF2BP-KO and WT mice.Original magnification, x400. H, Thelevels of HSFs (heat shock factors) and HSPs (heat shock proteins) in liver ofHSF2BP-KO and WT mice. I, The levelsof inflammatory factors in liver of HSF2BP-KO and WT mice. Results areexpressed as mean ± SE (n = 4-6/group) and compared by t-test or one-way ANOVA.* p &lt; 0.05. Figure S5. HSF2BP isupregulated in hepatic I/R injury. Liver immunohistochemical staining of HSF2BP (A) and its IHCscore (B)in liver transplant patients and healthy individuals. Western blot analysis ofHSF2BP (C) and its quantitativeresult (D) in human livers of livertransplant patients (n = 4) and healthy individuals (n = 3). Western blotanalysis of HSF2BP (E) and itsquantitative result (F) in mouselivers subjected to ischemia and reperfusion (n = 4-6/group). Western blotanalysis of HSF2BP (G) and itsquantitative result (H) in culturedhepatocytes under H/R condition (n = 3/group). Results are expressed as mean ±SE and compared by t-test or one-way ANOVA. * p &lt; 0.05 versus sham mice orcontrol group. Figure S6. CDC73 expressionlevels are increased in liver transplant patients and hepatic I/R mice.Western blot analysis ofCDC73 (A) and its quantitativeanalysis (B) in livers of livertransplant patients (n = 3) and healthy individuals (n = 4). Results areexpressed as mean ± SE and compared by t-test. * p &lt; 0.05. Western blotanalysis of CDC73 (C) and itsquantitative analysis (D) in mouselivers. Results are expressed as mean ± SE (n = 4) and compared by one-wayANOVA. * p &lt; 0.05 versus sham mice, # p &lt; 0.05 versus NTG mice. Figure S7. HSF2BP expression is increased after transfection with HSF2BPoverexpressed lentivirus in cultured hepatocytes. Western blot analysis ofHSF2BP (A) and its quantitativeanalysis (B) after transfection withLv-HSF2BP in HL-7702 cells. C, The HSF2BP mRNA level after transfection withLv-HSF2BP in HL-7702 cells. Results are expressed as mean ± SE (n = 3-5/group)and compared by t-test. * p &lt; 0.05. Figure S8. CDC73 expression is knocked down after transfection with siRNA-CDC73 in primaryhepatocytes. Westernblot analysis of CDC73 (A) and itsquantitative analysis (B) aftertransfection with siRNA-CDC73 in primary hepatocytes. Results are expressed asmean ± SE (n = 4/group) and compared by one-way ANOVA. * p &lt; 0.05 versussiRNA-NC group, # p &lt; 0.05 versus NTG-H/R group. Figure S9. Hepatic ischemicpreconditioning (HIPC) up-regulated the expression of HSF2BP, and HSF2BPknockout eliminated HIPC’s hepatoprotective effect during hepatic I/R. HIPC was induced by liverischemia for 10 min, followed by 10 min of reperfusion. Western blot analysisof HSF2BP (A) and its quantitativeanalysis (B) after HIPC. Results areexpressed as mean ± SE (n = 4-6/group) and compared by t-test. * p &lt; 0.05versus sham group. C, Liver H&amp;Estaining. Original magnification, x100 and x200. Liver necrosis area (D) and histological score (E). Results are expressed as mean ± SE(n = 4/group) and compared by one-way ANOVA. * p &lt; 0.05 versus WT mice, # p&lt; 0.05 versus I/R group.