Journal: Nature communications

Article Title: Targeting endogenous kidney regeneration using anti-IL11 therapy in acute and chronic models of kidney disease.

doi: 10.1038/s41467-022-35306-1

Figure Lengend Snippet: Fig. 1 | Autocrine IL11 activity initiates a pro-inflammatory and mesenchymal program in tubular epithelial cells via activation of both STAT3 and ERK. a Representative immunofluorescence (IF) images (scale bars: 100 µm) of EGFP and E-Cadherin expression in the kidneys of Il11EGFP/+ mice following folic acid (FA) injection (kidneys were collected atday 3, day21, and day 28, representative dataset from n = 3/group. b Representative IF images of IL11RA staining of primary human renal TECs (scale bars: 100 µm; representative dataset from n = 3/group). c ELISA of IL11 secretion from TGFβ1-stimulated TECs (24 h, n = 4/group). d Western blots of STAT3, MEK, ERK, p90RSK, and GSK3β activation and expression levels of SNAI1 and E-Cadherin in IL11-stimulated TECs over a time course (representative dataset from n = 4/group). e, f Data for RNA sequencing (RNA-seq) experiments on TEC stimulated with IL11 for 0 (Bsl), 1, 6, and 24 h. e Principal component analysis (PCA) of RNA-seq across the time-series. PC1 and PC2 account for 59% and 18% variance of gene expression, respectively. f Volcano plot displaying the adjusted p-value (−log10(p-adj)) and fold change (log2(Fold change)) of genes between IL11-

Article Snippet: For replication, and to exclude potential off-target effects, we repeated the preemptive treatment experiment with a second, commercially available, neutralizing IL11 antibody (MAB218, R&D Systems) and compared its effects to X203 (Fig. 3d).

Techniques: Activity Assay, Activation Assay, Expressing, Injection, Staining, Enzyme-linked Immunosorbent Assay, Western Blot, RNA Sequencing, Gene Expression

Journal: Nature communications

Article Title: Targeting endogenous kidney regeneration using anti-IL11 therapy in acute and chronic models of kidney disease.

doi: 10.1038/s41467-022-35306-1

Figure Lengend Snippet: Fig. 5 | Therapeutic targeting of IL11 protects mice from UUO-induced renal tubule damage, inflammation, and fibrosis. a Representative immuno- fluorescence images (scale bars: 100 µm) of EGFP and E-Cadherin expression in UUO and control kidneys of Il11EGFP/+ mice (representative dataset from n = 3/group) and b Western blots of IL11 protein expression in control and UUO kidneys (representative dataset from n = 5/group). Kidneys were collected at day 10 and contralateral kidneys were used as controls. c Schematic of X203 therapeutic dosing of mice subjected to left unilateral ureteral obstruction for experiments shown in (d–h). IgG/X203 (10 mg/kg) was administered on day 4 and day 8 post UUO and kidneys were collected on day 10; contralateral (right) kidneys from the IgG group were used as controls. d Western blots of pERK, ERK, pp90RSK, p90RSK, pGSK3β, GSK3β, SNAI1, ZEB, E-Cadherin, Cyclin D1, αSMA, FN1, and GAPDH

Article Snippet: For replication, and to exclude potential off-target effects, we repeated the preemptive treatment experiment with a second, commercially available, neutralizing IL11 antibody (MAB218, R&D Systems) and compared its effects to X203 (Fig. 3d).

Techniques: Expressing, Control, Western Blot

Journal: Nature communications

Article Title: Targeting endogenous kidney regeneration using anti-IL11 therapy in acute and chronic models of kidney disease.

doi: 10.1038/s41467-022-35306-1

Figure Lengend Snippet: Fig. 9 | Anti-IL11 promotes kidney regeneration and reverses renal dysfunction in chronic kidney disease. a Schematic of X203 therapeutic dosing regimen for CKD reversal experiments shown in (b–l): X203 or IgG (IP, 10 mg/kg) was admi- nistered 2X/week for 12 weeks from day 21 (week 3) after FA injury, when CKD is established and sustained. Blood and kidneys were collected every 3 weeks; full course serum and kidney phenotyping study were performed at the study end- point (week 15: 12 weeks after the start of X203 therapy); blood and kidneys from control mice i.e., those receiving IP injection of vehicle control (0.3 M NaHCO3) were collected at week 3 and 15. b Representative kidney gross anatomy, c kidney weights (NaHCO3 (W3), FA (W3) (n = 11/group), FA + IgG (W6, W9), FA + X203 (W6, W12) (n = 4/group), FA + X203 (W9), FA + IgG (W12), NaHCO3 (W15) (n = 5/group), FA + IgG/X203 (W15) n = 13/group)), d renal collagen content by hydroxyproline assay, e representative Masson’s Trichrome images of whole kidney cross section (scale bars: 500 µm, representative dataset from n = 4/group for kidneys collected at week 6, 9, 12 or from n = 5/group for week 3 and 15), f Western blots showing renal ERK and p90RSK activation, GSK3β inactivation, and SNAI1, ZEB, E- Cadherin, Cyclin D1, αSMA, FN1, GAPDH expression (representative dataset from

Article Snippet: For replication, and to exclude potential off-target effects, we repeated the preemptive treatment experiment with a second, commercially available, neutralizing IL11 antibody (MAB218, R&D Systems) and compared its effects to X203 (Fig. 3d).

Techniques: Control, Injection, Hydroxyproline Assay, Western Blot, Activation Assay, Expressing