In BMP4-stimulated C2C12BRA reporter cells, DMH1 reduced Id1-promoter luciferase activity, with an approximately 100 nM IC50 reported by the authors. This cellular reporter endpoint differs from purified-enzyme potency. [1]
During seven-day neural induction with 10 µM SB431542, increasing DMH1 increased SOX1-positive cells in human iPSC cultures while PAX6 expression changed little across the tested concentrations. The authors reported comparable SOX1-positive cell frequency at 5 µM DMH1 and 500 ng/mL Noggin, but substantial toxicity at 10 µM DMH1. [2]
In HeLa and MCF-7 cells, 5–10 µmol/L DMH1 reduced the 5-FU-induced LC3-II/LC3-I response. However, the authors reported no enhancement of 5-FU-mediated viability inhibition at 24, 48 or 72 hours. The LC3 readout alone does not establish autophagic flux. [3]
Purified kinase inhibition and selectivity panel
Biochemical assay · Purified human ALK2, ALK5, AMPK, KDR and PDGFRβ [1]
DMH1 directly inhibited ALK2 in the reported kinase panel. Staurosporine served as a nonspecific kinase-inhibitor control. The four other tested kinases were reported as showing no inhibition under the assay conditions; this is a limited panel, not kinome-wide selectivity.
Literature-Reported Treatment Conditions
- Treatment concentration
- Ten concentrations by 3-fold serial dilution starting at 30 µM
- Method
- Radiometric kinase assay with 10 µM (33P)γATP
- Reported limits
- Reaction duration, assay vehicle, replicate count and IC50 uncertainty are not specified in the selected kinase methods/table.
Methods and source locations
Compound reported in the paper: DMH1; Compound label in the paper: DMH-1
Evidence for the reported research material and model; does not establish equivalence of an Adooq lot or a salt formulation.
Table 2 and footnote; Methods, Kinase assay; Figure 3 and caption
Purified human ALK2 kinase activity: Radiometric kinase assay; 10 µM ATP — Table 2, DMH1 row, ALK2 column; Methods, Kinase assay
ALK5, AMPK, KDR and PDGFRβ inhibition: Same panel; maximum tested compound concentration 30 µM. Table defines “No” as no inhibition. — Table 2, DMH1 row and footnote; Methods, Kinase assay
BMP4-responsive transcriptional reporter inhibition
Cell-based study · Mouse C2C12BRA cells carrying the Id1 promoter–firefly luciferase reporter [1]
DMH1 reduced the BMP4-induced reporter signal in a concentration-dependent manner. The experiment included unstimulated and BMP4-only controls, and luciferase results were normalized to cell titers.
Literature-Reported Treatment Conditions
- Treatment concentration
- Figure 4b: 0, 0.05, 0.1, 0.2 and 0.5 µM DMH1
- Exposure duration
- Overnight
- Method
- BMP-responsive luciferase assay normalized using Cell Titer-Glo
- Co-treatment
- BMP4, 50 ng/mL
- Reported limits
- Reporter assay vehicle, exact overnight duration, biological replicate count and error-bar definition are not specified in the selected caption/methods.
Methods and source locations
Compound reported in the paper: DMH1; Compound label in the paper: DMH-1
Evidence for the reported research material and model; does not establish equivalence of an Adooq lot or a salt formulation.
Figure 4b and caption; Results paragraph beginning “Next, we examined”; Methods, BMP-responsive luciferase reporter assays; Figure 3
BMP4-induced Id1-promoter luciferase signal: Cellular reporter assay; this is not a purified-enzyme IC50 or a treatment recommendation. — Results paragraph beginning “Next, we examined”; Figure 4b; Methods, BMP-responsive luciferase reporter assays
Concentration-dependent neural precursor marker response
Cell-based study · Human induced pluripotent stem cell lines SM3 and CA6; SM3 immunocytochemistry and both lines by flow cytometry [2]
During dual-SMAD neural induction, increasing DMH1 increased SOX1-positive cells while PAX6 expression was comparatively insensitive to the concentration series. At 5 µM, SOX1-positive cell frequency was reported as comparable to Noggin (500 ng/mL). At 10 µM, the authors reported substantial toxicity with lower cell density. These are combination conditions with SB431542.
Literature-Reported Treatment Conditions
- Treatment concentration
- DMH1 0.5, 2, 5 or 10 µM
- Exposure duration
- 7 days
- Method
- Neural induction; day-7 PAX6/SOX1 immunocytochemistry and flow cytometry
- Co-treatment
- SB431542 10 µM in neural induction conditions; comparator: Noggin 500 ng/mL with SB431542; mTeSR maintenance-medium control
- Reported limits
- Vehicle and independent replicate count for Figure 4 are not specified there. Flow methods acquired 10,000 events/sample; events are not independent biological replicates.
Methods and source locations
Compound reported in the paper: DMH1; Compound label in the paper: DMH-1
Evidence for the reported research material and model; does not establish equivalence of an Adooq lot or a salt formulation.
Figure 4a–b and caption; Results paragraphs surrounding Figure 4; Methods, Neural Induction and Flow Cytometry
SOX1- and PAX6-positive cell frequency at day 7: Immunocytochemistry and flow cytometry; no exact percentages transcribed from graph. — Figure 4a–b; Results paragraphs immediately following Figure 4
Cell density/toxicity during neural induction: Author-described observation with day-7 microscopy; no numerical toxicity fraction reported in selected text. — Results paragraph immediately following Figure 4; Figure 4a
5-FU combination: LC3 response without enhanced viability inhibition
Cell-based study · Human HeLa cervical cancer and MCF-7 breast cancer cells [3]
DMH1 reduced the 5-FU-induced LC3-II/LC3-I ratio in both cell lines, but the authors reported no enhancement of 5-FU-mediated viability inhibition at 24, 48 or 72 hours. DMH1-only groups were included. Reduced LC3 ratios alone do not establish autophagic flux or a causal BMP mechanism.
Literature-Reported Treatment Conditions
- Treatment concentration
- DMH1 5 or 10 µmol/L; 5-FU 75 µmol/L for LC3 blots and 7.5–300 µmol/L for the viability concentration series
- Exposure duration
- LC3 assays: 24 h; viability assays: 24, 48 and 72 h
- Method
- Western blotting and MTT assay
- Co-treatment
- 5-fluorouracil (5-FU); untreated, DMH1-only and 5-FU-only controls
- Reported limits
- Vehicle concentration is not specified for this experiment. n labels are retained as reported without assuming independent biological replicates.
Methods and source locations
Compound reported in the paper: DMH1; Compound label in the paper: DMH-1
Evidence for the reported research material and model; does not establish equivalence of an Adooq lot or a salt formulation.
Figure 4a–d and caption; Results 3.4; Methods 2.1–2.3, 2.6 and 2.8
5-FU-induced LC3-II/LC3-I ratio: Actin-controlled Western blot; mean±SEM per Methods 2.8. Figure 4a/c show comparisons to 5-FU alone. — Figure 4a,c (image labels and caption); Results 3.4; Methods 2.8
Viability response to 5-FU with versus without DMH1: MTT; Figure 4b/d labels NS versus counterpart drug dose; mean±SEM. n experimental unit is not specified. — Figure 4b,d and caption; Results 3.4; Methods 2.3 and 2.8
