Journal: Signal Transduction and Targeted Therapy

Article Title: Analogous comparison unravels heightened antiviral defense and boosted viral infection upon immunosuppression in bat organoids

doi: 10.1038/s41392-022-01247-w

Figure Lengend Snippet: Bat organoids elicited a more robust antiviral response to Poly(I:C) treatment. Bat and human intestinal organoids were sheared mechanically and treated with 10 μg/ml Poly(I:C) or mock-treated with DMSO. a , b Induction of IFNs (left) and ISGs (right) in bat intestinal organoids ( a ) and human intestinal organoids ( b ) at the indicated hours after Poly(I:C) treatment. Results show the log2-fold change of GAPDH-normalized expression level in the treated organoids relative to mock-treated organoids. Data represent the mean and s.d. of a representative experiment in organoids from a bat and human donor, n = 3. c Culture media from the treated or mock-treated human intestinal organoids at the indicated hours were applied to ELISA to measure concentrations of IFNL1 and IFNL3. Data represent the mean and s.d. of a representative experiment in one line of human organoids, n = 3. Two-tailed unpaired Student’s t test. d Poly(I:C) treated or mock-treated human intestinal organoids were collected at the indicated hours post-treatment and subjected to Western blot to detect human ISG15. e , f Bat and human intestinal organoids were sheared and incubated with 10 μg/ml Poly(I:C) Fluorescein in triplicate for 2 h and 6 h. The organoids were then dissociated and applied to flow cytometry to detect the percentage at 2 h post treatment ( e ) and mean fluorescence intensity (MFI, f ) of Fluorescein-positive cells at 2 and 6 h post treatment. g , h Induction of TNF-a and IL6 ( g ) and IP10 ( h ) in bat and human intestinal organoids at the indicated hours after treatment. Results show the fold change of GAPDH-normalized expression level in Poly(I:C)-treated organoids relative to mock-treated organoids. Data represent the mean and s.d. of a representative experiment in organoids from a bat and human donor, n = 3

Article Snippet: Cell-free media were harvested from treated and mock-treated human intestinal organoids for measuring the amount of human IFNL1 and IFNL3 using ELISA kits (R&D Systems, DY7246, D28B00).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Western Blot, Incubation, Flow Cytometry, Fluorescence

Journal: Signal Transduction and Targeted Therapy

Article Title: Analogous comparison unravels heightened antiviral defense and boosted viral infection upon immunosuppression in bat organoids

doi: 10.1038/s41392-022-01247-w

Figure Lengend Snippet: TLR3 and RLR signaling pathways mediate antiviral responses in bat and human organoids. a A schematic graph outlines the experimental procedure for panels b – g . Bat and human intestinal organoids were pretreated with BX795 (0, 0.1 and 1 μM) or CYT387 (0, 0.1, and 1 μg/ml) overnight. After mechanical shearing, the organoids were incubated with or without 10 μg/ml Poly(I:C), together with the initial concentrations of BX795 or CYT387. At the indicated hours after Poly(I:C) treatment, the organoids were harvested and subjected to RT-qPCR assay to examine mRNA expression levels of bat and human IFNL1 and IFNL3; cell-free media from human organoids were applied to ELISA to detect IFNL1 and IFNL3. Data represent the mean and s.d. of a representative experiment in organoids from a bat and human donor, n = 3. Ordinary one-way ANOVA with Tukey’s multiple comparison test. b , c Induction of IFNL1 (left) and IFNL3 (right) in bat intestinal organoids treated with BX795 ( b ) and CYT387 ( c ). Results show the fold change of GAPDH-normalized expression level in the Poly(I:C)-treated organoids relative to mock-treated organoids. d , e Induction of IFNL1 and IFNL3 in human intestinal organoids treated with BX795 ( d ) and CYT387 ( e ). IFNL1 ( f ) and IFNL3 ( g ) secretion from the human organoids at 4 h post-stimulation. h A schematic graph describes the experimental procedure for panels i and j . Bat and human intestinal organoids were pretreated with 1 μg/ml CYT387 or DMSO overnight. After mechanical shearing, the organoids were incubated with 1 μg/ml CYT387 or DMSO with or without 10 μg/ml Poly(I:C) for 4 h. Cell-free media were then harvested and subjected to PRM-MS to analyze bat IFNL3 ( i ) and ISG15 of bat and human ( j ). The dotted lines represent the detection limit. Data represent the mean and s.d. of a representative experiment in organoids from a bat and human donor, n = 3. Two-tailed unpaired Student’s t test

Article Snippet: Cell-free media were harvested from treated and mock-treated human intestinal organoids for measuring the amount of human IFNL1 and IFNL3 using ELISA kits (R&D Systems, DY7246, D28B00).

Techniques: Protein-Protein interactions, Incubation, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Comparison, Two Tailed Test

Journal: Blood Cancer Journal

Article Title: Anti-tumor activity against multiple myeloma by combination of KW-2478, an Hsp90 inhibitor, with bortezomib

doi: 10.1038/bcj.2012.13

Figure Lengend Snippet: Anti-tumor activity in OPM-2/GFP orthotopic model. OPM-2/GFP cells (1 × 10 7 cells/mouse) were inoculated i.v. into severe combined immunodeficient (SCID) mice. ( a ) Thirty-three days after cell inoculation, GFP fluorescence was measured followed by histochmical examination, ( b ) Twenty-seven days after cell inoculation, KW-2478 (KW) or/and bortezomib (BTZ) were administered i.v. twice weekly for 2 weeks. Photographs were taken 50 days after cell inoculation from dorsal angle. ( c ) Forty days after cell inoculation, serum M protein concentration was measured by ELISA. Each column represents the mean+s.d. n =5. * P <0.05; Tukey's test.

Article Snippet: On day 26, serum M protein (Igλ chain) of each mouse was measured using the Human Lambda ELISA Quantitation Kit (Bethyl Laboratories).

Techniques: Activity Assay, Fluorescence, Protein Concentration, Enzyme-linked Immunosorbent Assay