The Hypothesis That Failed (And Why That's Valuable)

The Question

David had been reading about Lion's Mane mushroom (Hericium erinaceus) and its supposed cognitive benefits. He'd seen claims that it promotes nerve growth factor (NGF) production.

He wondered: does the Lion's Mane supplement he actually buys do anything measurable?

The Hypothesis

David's initial idea: "My Lion's Mane supplement boosts brain function."

This is untestable on Litmus (no human subjects), so he refined it to something mechanistic:

Final hypothesis: "Hot water extract from my Lion's Mane supplement (Brand X, 500mg capsules) induces NGF secretion in PC12 cells at ≥2x baseline levels at 100μg/mL concentration."

Rationale: If the supplement works via NGF induction (as claimed), extract from the actual capsules should show measurable activity in a standard NGF-responsive cell line.

The Results

NGF ELISA Results

Cell Viability (MTT)

The Verdict

Hypothesis rejected.

No concentration showed statistically significant NGF induction (all p > 0.05). The highest fold-change observed was 1.24x, well below the 2x threshold. Cell viability was fine, so this isn't a toxicity artifact.

The positive control worked perfectly (39x induction), confirming the assay itself was functional.

What David Learned

1. The Supplement May Not Contain Active Compounds

Lion's Mane's purported activity comes from compounds called hericenones and erinacines. These require specific extraction methods. A commercial supplement might:

• Use cultivation methods that don't produce these compounds
• Use mycelium-on-grain (mostly starch) rather than fruiting bodies
• Undergo processing that degrades active compounds

2. The Claim May Be Overstated

Even studies showing Lion's Mane activity often use:

• Purified compounds, not crude extracts
• Higher concentrations
• Different cell types

The marketing claim that a 500mg capsule "supports NGF" is several steps removed from controlled research.

3. Negative Results Are Data

David didn't get the result he hoped for, but he got an answer: his specific supplement, tested in a standard assay, doesn't show detectable NGF induction.

This is exactly what science is supposed to do—test claims and accept the results.

Was It Worth $400?

David's take:

"I've probably spent more than $400 on Lion's Mane supplements over the years based on vague promises. Now I know that at least this brand, in this assay, doesn't do what the marketing implies.

I could buy a different brand that claims to use fruiting bodies and specific extraction methods. Or I could accept that the evidence isn't there and spend my money elsewhere.

Either way, I made a decision based on data instead of marketing."

Why Open Results Matter

David published his results as open access. Now when someone searches "Lion's Mane NGF Brand X," they'll find actual data rather than just marketing claims.

Three months after publication:

• 847 views
• 12 saves
• 3 citations in forum discussions

Lessons for Citizen Scientists

1. Negative results are still results: Rejecting a hypothesis is a valid scientific outcome

2. Test what you actually use: Studies on purified compounds don't tell you about your specific supplement

3. Pre-register your criteria: David defined "success" as ≥2x induction before seeing data. This prevented retroactive goalpost moving.

4. Consider what you'll do with each outcome: Before submitting, think through: "If positive, then what? If negative, then what?"

5. Share openly: Your negative result might save someone else $400 and months of false hope