Journal: PLoS ONE

Article Title: Differential Expression of Complement Markers in Normal and AMD Transmitochondrial Cybrids

doi: 10.1371/journal.pone.0159828

Figure Lengend Snippet: Complement Proteins’ Western Blotting Information.

Article Snippet: 6 , CFB , 85 kDa , Goat Anti-factor B Polyclonal antibody # AF2739 (RD Systems) , 1:1000 , Human , Dnk pAb to Goat IgG (HRP) ab97120 (ABCAM) , beta-actin antibody GTX 110564 (Genetex).

Techniques: Western Blot

Journal: Cell death & disease

Article Title: Mechanistic target of rapamycin complex 1 orchestrates the interplay between hepatocytes and Kupffer cells to determine the outcome of immune-mediated hepatitis.

doi: 10.1038/s41419-022-05487-0

Figure Lengend Snippet: Fig. 5 The activation of mTORC1 in KCs enhances complement alternative system. a Expression heatmap of genes of neutrophils chemotaxis or complement activation analyzed by RNA-seq from Tsc1+/+ and Tsc1-/- BMMs (n = 3 each). b Western blotting result was shown the expression of CFB protein in hepatic tissues from mice. c Left, representative co-immunofluorescent staining images for F4/80 with CFB. Scale bar = 50 μm. Right, quantitative determination of F4/80+ and CFB+ cells among groups as indicated, n = 3. d Western blotting assay showing the abundance for TSC1, CFB, and p-S6 in BMMs. e qRT-PCR analysis showing the CFB mRNA abundance in BMMs, n = 3. f Western blotting assay showing the abundance for TSC1, CFB, and p-S6 in KCs. g qRT-PCR analysis showing the CFB mRNA abundance in KCs, n = 3. h Representative immunofluorescent staining images for C3d. Scale bar = 50 μm. i Representative immunostaining images for C5b-9. Scale bar = 50 μm. j Western blotting assay showing the abundance for Rheb and CFB in BMMs. k Western blotting assay showing the abundance for Rheb and CFB in KCs. l Representative co-immunofluorescent staining images for F4/80 with CFB (white arrows). Scale bar = 100 μm. m Quantitative determination of F4/80+ and CFB+ cells among groups as indicated, n = 3. n Representative immunofluorescent staining images for C3d. Scale bar = 100 μm. o Representative immunostaining images for C5b-9 among groups as indicated. Scale bar = 100 μm. *p < 0.05.

Article Snippet: Three μm sections were cut and immunolabelled with primary antibodies specifically binding F4/80 (cat: 14–4801-82, Invitrogen, USA), p-S6 (Ser235/236) (cat: 4858, Cell Signaling Technology, USA), CD11b (cat: 557396, BD Biosciences), CD3 (cat: 555273, BD Biosciences), ly6b (cat: MCA771G, Bio-Rad, California, USA), Ki67(cat: ab15580, Abcam, Cambridge, UK), cleaved Caspase 3 (cat: 9664, Cell Signaling Technology, USA), CFB (cat: NBP1-89985, Novus; cat: ab192577, Abcam, Cambridge, UK), C3d (cat: AF2655, R&D Systems) or TdT-mediated dUTP-biotin nick end labeling (TUNEL, Promega, Madison, WI).

Techniques: Activation Assay, Expressing, Chemotaxis Assay, RNA Sequencing, Western Blot, Staining, Quantitative RT-PCR, Immunostaining

Journal: Cell death & disease

Article Title: Mechanistic target of rapamycin complex 1 orchestrates the interplay between hepatocytes and Kupffer cells to determine the outcome of immune-mediated hepatitis.

doi: 10.1038/s41419-022-05487-0

Figure Lengend Snippet: Fig. 6 Down regulation of CFB in liver protects against Con-A induced liver injury. a Western blotting assay showing CFB expression in mouse livers after shRNA-CFB injection. b Representative co-immunofluorescent staining images for F4/80 with CFB (white arrows). Scale bar = 100 μm. c The strategy for establishing a mouse model of injection of shRNA-CFB and Con-A administration. d The ALT levels in serum of mice exposed to Con-A for 8 h, n = 6. e Representative HE-stained mouse livers. Scale bar = 100 μm. f Liver sections of were immunofluorescent stained with TUNEL. Scale bar = 200 μm. g Left, representative immunofluorescent staining images for ly6b. Scale bar = 100 μm. Right, quantitative determination of ly6b+ cells among groups as indicated, n = 4. h Left, representative immunofluorescent staining images for C3d. Scale bar = 100 μm. Right, quantitative determination of C3d area in a field of vision, n = 4. i Representative immunostaining images for C5b-9. Scale bar = 100 μm. j Left, representative immunofluorescent staining images for F4/80 (white arrows). Scale bar = 100 μm. Right, quantitative determination of F4/80+ cells among groups as indicated, n = 4. *p < 0.05. k Schematic working model on the role of mTORC1 activation in hepatocytes and KCs in the pathogenesis of ALD.

Article Snippet: Three μm sections were cut and immunolabelled with primary antibodies specifically binding F4/80 (cat: 14–4801-82, Invitrogen, USA), p-S6 (Ser235/236) (cat: 4858, Cell Signaling Technology, USA), CD11b (cat: 557396, BD Biosciences), CD3 (cat: 555273, BD Biosciences), ly6b (cat: MCA771G, Bio-Rad, California, USA), Ki67(cat: ab15580, Abcam, Cambridge, UK), cleaved Caspase 3 (cat: 9664, Cell Signaling Technology, USA), CFB (cat: NBP1-89985, Novus; cat: ab192577, Abcam, Cambridge, UK), C3d (cat: AF2655, R&D Systems) or TdT-mediated dUTP-biotin nick end labeling (TUNEL, Promega, Madison, WI).

Techniques: Western Blot, Expressing, shRNA, Injection, Staining, TUNEL Assay, Immunostaining, Activation Assay